Substituted thiazolidine-4-ones

ABSTRACT

HIGHLY ACTIVE ANTIBACTERIALS ARE DESCRIBED WHICH ARE 5SUBSTITUTED THIAZOLIDINE-4-ONES HAVING FORMULA   2,4-DI(O=),3-(X-CH2-CH2-),5-((5-(O2N-)FUR-2-YL)-CH=)   THIAZOLIDINE   IN WHICH X IS CL OR BR. TESTS ARE DESCRIBED IN WHICH ACTIVITY AGAINST A LARGE NUMBER OF GRAM POSITIVE AND GRAM NEGATIVE BACTERIA ARE XXXXTED.

United States Patent Int. Cl. C07d 91/14 US. Cl. 260-240 A 3 Claims ABSTRACT OF THE DISCLOSURE Highly active antibacterials are described which are S- substituted thiazolidine-4-ones having formula in which X is C1 or Br. Tests are described in which activity against a large number of gram positive and gram negative bacteria are reported.

This application is a continuation-in-part of application, Ser. No. 733,808, filed June 3, 1968, now US. Pat. 3,678,041 granted July 18, 1972.

This invention relates to a new series of chemical compounds which have a high antibacterial activity against a large number of gram positive and gram negative bacteria and which also have significant antitubercular activity; they are also capable of inhibiting the growth of a large number of pathogenic fungi such as candida, epidermophyton and trichophyton. By reason of these properties, the products are therapeutically useful both in human medicine and in the veterinary field.

According to the present invention there is provided a S-hubstituted thiazolidine-4-one having one of the general formulae in which n is 0 or 1, R is a hydrogen atom or a bromine atom, R is a hydrogen atom, an alkyl, or alkenyl group which may be substituted by a halogen atom, or a hydroxyl, a carboxylic acid, carboxarnide, substituted carboxhydrazide or carboxylic acid ester group or a primary, secondary or tertiary amino group, or R is a carbalkoxy group, an alkynyl group, an azomethine group or a cycloalkyl, aryl or aralkyl group which cycloalkyl, aryl or aralkyl group may be substituted by one or more halogen atoms or hydroxyl, sulphonamido, nitro or carboxylic acid groups, X is an oxygen or sulphur atom or an imino group having the formula NR in which R is a hydrogen atom or an alkyl, alkenyl, cycloalkyl, aryl, aralkyl or arylsulphonyl group which groups may be substituted,

or a carbamido or amidinomethyl group and each of R and R is a hydrogen atom, or an alkyl, alkenyl, cycloalkyl, aryl or aralkyl group, which group may be substituted, or R and R together with the nitrogen atom to which they are attached form a heterocyclic ring, and salts of such thiazolidine-4-ones with acids and bases.

3,704,296 Patented Nov. 28, 1972 The compounds having the above general formula may contain a free carboxyl group in which case they form salts with mineral and organic bases such as the alkali metal bases, ammonia and amines. On the other hand they may contain primary, secondary and tertiary amino groups in which case they form salts with mineral and organic acids, especially with pharmaceutically acceptable mineral and organic acids, e.g. the hydrochlorides.

According to a feature of the invention the S-substituted thiazolidine-4-ones having the above general formulae may be produced by a process which comprises reacting a S-nitrofuran aldehyde having the general forin which n and R are as above defined, with a thiazolidine- 4-one having one of the general formulae in which X, R R and R are as above defined, in the presence of a dehydrating agent.

The S-nitrofuran aldehyde used is S-nitofurfural, )8- (5-nitrofuryl-2)acrolein or a-bromo-p-(5-nitrofuryl-2)- acrolein.

The thiazolidine-4-one employed may or may not have a substituent present in the 3-position. When there is no substituent in the 3-position the condensation product, which corresponds to the following formula:

R 11 (III) may be reacted, with one or more compounds which act as a source of the substituent R As an example formaldehyde may be used for introducing a substituent R When it is desired to produce a compound which liberates formaldehyde, this may be used alone with a view to hydroxymethylation, or together with a secondary amine when it is desired to carry out a Mannich reaction. Alternatively, one may start with a halogenated derivative R Hal, the symbol Hal denoting a halogen atom, and react this with an alkali metal derivative of the Compound III.

Rhodanine which has the following formula as well as most of its derivatives corresponding to the general formula s (IV) in which R is as defined above other than hydrogen, are already known. The remaining derivatives IV may be obtained by condensation of monochloroacetic acid with an alkali metal dithiocarbamate and subsequent closure of the lactam ring by heating in the presence of a strong mineral acid.

Condensation of a rhodanine IV with the nitrofuran aldehyde may be carried out in the presence of a dehydrating agent such as anhydrous sodium or potassium acetate in a single diluent (for example methanol, ethanol or acetic acid) or, more preferably, in a mixture of diluents (for example, a mixture of ethanol and acetic acid). The condensation is carried out by heating the reactants to between 60 and 100 C. for a time which is usually between 1 and 4 hours. The resulting products generally precipitate; they are isolated after cooling by the usual methods and may be recrystallised from solvents such as nitromethane and dimethylformamide.

If compounds having the general Formula III in which R=H, n= or 1 and X=S are treated with formaldehyde or trioxymethylene in solution in dimethylformamide and subjected to moderate heating, the corresponding hydroxymethyl derivatives I (R =CHOH) are obtained in good yields. The presence of a catalyst is not essential although traces of organic bases such as piperidine ormorpholine may be added. The bond connecting the hydroxymethyl group to the nuclear nitrogen atom is not very stable and breaks on heating above 120 C., liberating the starting material. By treating compounds III (X=S) with the stoichimetrical quantities of formaldehyde (or trioxymethylene) and a secondary amine, for example, morpholine in suspension in ethanol or in solution in dimethyl formamide, the corresponding Mannich base is formed even at room temperature.

Appropriately substituted thiazoli-dine-4-ones may also be obtained by treating rhodanine N-acetic acid (R ='CH COOH in Formula IV) with thionyl chloride in solution in benzene and then reacting the resulting acid chloride with ammonia or with a secondary amine such as pyrrolidine or morpholine or with a hydrazide such as isonicotinic acid hydrazide.

S-amino-rhodanine (R =NH in Formula IV) undergoes reaction with two molecules of the nitrofuran aldehyde, one reaction taking place at the 3-position with the formation of the hydrazone in a mixture of ethanol and mineral acid, whilst the other takes place at the 5-position by reason.of an aldolisation reaction in acetic acid in the presence of an anhydrous alkali metal acetate.

The starting compound thiazolidine-4-one may also be a derivative of thiazolidine2,4-dione In this case, if R is a substituent other than a hydrogen atom, this substituent may have been introduced into the thiazolidine-2,4dione by means of one of its alkali metal salts, generally the potassium salt formed by treatment with potash in solution in ethanol at room temperature (the corresponding sodium salt is obtained in analogous manner or by means of sodium hydride). This salt is condensed with a halogen compound R Hal in which Hal is a halogen atom, preferably bromine or an iodine, in solution either in methanol or in ethanol under refiux or in dimethyl formamide under reflux if these halogens are relatively non-volatile. Derivatives in which R is an alkylgroup can be obtained by means of other alkylating agents, such as by the action of diazomethane (if R =CH or by reaction with dialkylsulphates.

Condensation of the nitrofuran aldehydes is conveniently carried out by heating in solution in acetic acid in the presence of anhydrous sodium or potassium acetate at temperatures in the region of 100 C. until no further precipitation of the condensation product takes place. After prolonged cooling, the solid is isolated and recrystallized from a solvent which may be chosen from the following: methanol, ethanol, acetic acid, 'dioxane and dimethylformamide. The following reaction scheme illustrates the series of reactions enumerated above:

LJO

If, in the final product, it is desired that R be other than hydrogen, it is sometimes preferable to condense a thiazolidine-2,4-dione which is unsubstituted in the 3-position with a nitrofuran aldehyde and to convert the product into an alkali metal derivative e.g. by means of an ethanol solution of sodium or potassium hydroxide at room temperature. The derivative thus obtained then reacts rapidly with a compound R Hal in which Hal is a halogen in solution in methanol under reflux or, even better, in dimethylformamide at about C. After concentration of the solvent under reduced pressure and treatment with water, the products are obtained in very good yields.

Another process for obtaining the same products comprises hydrolysing the corresponding 2-imino-derivatives containing the substituent R in the 3-position with a strong mineral acid for several hours at about 100 C. according to the following equation:

KOH

This method, however, does not always give satisfactory results because a substantial proportion of unreacted starting material may be present in the product.

As in the rhodanine series, the condensation products in which the 3-position is free are capable of yielding Mannich bases with aliphatic secondary amines such as dimethylamine, diethylamine and diethanolamine or cyclic amines such as pyrrolidine, piperidine, morpholine, piperazine or 4-(fi-hydroxy-ethyl)piperazine under the same conditions as their homologues. Suitable diluents are ethanol (the reaction taking place in a heterogeneous medium), dioxane or dimethylformamide. Treatment with formaldehyde also leads to the formation of derivatives which are hydroxymethylated in the 3-position, these products having the same instability when heated as the homologous derivatives of rhodanine; in effect, they lose one equivalent of formaldehyde at temperatures from C. upwards.

The derivatives of 2-imino-thiazolide-4-one or pseudo thiohydanation (R =R =H in Formula VI) =NR= s v1 have the same characteristic feature as the homologues which have just been described, namely nucleophilic reactivity of the methylene group in the 5-position, as a i'esult of which they readily undergo the Kuoevenagel reaction with the S-nitrofuran aldehydes. 7

Compounds of the Formula VI above are generally Starting from unsymmetrically disubstituted thioureas, only the following type of amine can be formed:

C-N obtained by the condensation of N,N -d1subst1tuted l R: thioureas with monochloroacetic acid or with its methyl 5 k or ethyl ester. If on the other hand, a monosubstituted thiourea is used a mixture of two isomers (VIa and VIb) 0) be obtalnedt and one of P VIb Compounds of the Formula VIc may be obtained by isomer) may m be the result f' an equlglbnun} treating the sodium or potassium salts of the monosubtween two tautomeric forms (VIb and VIb as illus- 1O stituted compounds 3 with an alkylating agent trated by the following reactlont or by treating rhodanine with a secondary amine (which may be a secondary cyclic amine). All these types of products are obtained by the ring closure of the corresponding thioureas by heating with monochloroacetic acid COOR t or with its methyl or ethyl esters in a dlluent such as H201 ethanol, 2-propanol, n-butanol, acetic acid or benzene. 0 Depending upon whether or not sodium or potassium acetate is added the cyclisation product is obtained either C=NR in the form of the base or in the form of the hydrochloride. 0CNR S The aryl Z-sulphonylamido-derivatives may be obtained by treating the corresponding pseudothiohydantoin with C=NH an arylsulphonyl chloride in pyridine. The semicarbazone OC N (VIbZ) of the 2-derivative is obtained by treating the corresponda) ing rhodanine with semicarbazide in known manner NHRl whilst the 2-guanidyl derivative may be obtained by ring closure of thioglycollic acid with cyanoguanidine in aqueous solution Whilst heating. Condensation of all these derivatives with the nitrofuran aldehydes may be carried out under the conditions already mentioned above, namein which R" is an alkyl group and R is as hereinbefore y heating 111 acetic acid medium in the Presence of d fin d, anhydrous sodium or potass1um acetate until the product The results which we have obtained agree with those has precipitated; isolation and purification y be effected which have already bee bli h d nd onfirm th as previously described. It is not essential to isolate the dominance of the 2-iminothiazolide-4-one form (VIb thiazolidine-4-one before condensation with the nitrowhen R is an aryl or an acyl group and the predominance furan aldehyde since these two reactions of forming the of the 2-amino-2-thiazoline-4-one form (VIb When R heterocycle and condensation of the heterocycle can take is an alkyl, a cycloalkyl or an aralkyl substituent. Methylplace simultaneously. When the 3-position is unsubstithiourea generally leads to comparable quantities of the tuted, the products are capable of undergoing certain retwo isomers V121 and Vlb If the heating time in the actions, such as those leading to the formation ofthe cyclisation step is prolonged, the Wet isomer (labile form) corresponding Mannich bases by reaction with formaldemay be converted into the stable VIb isomer which has hyde and a secondary amine. been isolated in high yields. Typical compounds produced according to the inven- The structure of the above products is confirmed by tion are listed in the following tables; the examples to spectrographic studies. which the first column refers follow the tables:

TABLE I l I W OgN CH==C CH=C o:

o l j M.P. Example Code n R R (deg) Name 1 10290 0 Ethyl 3-ethy1-5-(5-nitr0furiurylidene)-2-thioxothlazolidine-4-one.

0 Methyl 190 3-methyl-5-(5-nltroiuriurylidene)-2-thioxothiazolidine-4-one. 0 Allyl 145 3-allyl-5-(6-nitroiurfurylidene)-2-thioxothlazolidinet-one. 0 n-Heptyl.-. 124 3-n-heptyl-5-(5-nltrofurfurylidene)-2-thioxothiazolidine-4-one. 0 n-Dodecyl 104 3-n-dodecyl-5-(5-nitrofurfurylidene)-2-thioxothiazolidinet-one. 0 n-HexadeeyL 114 3-n-hexadeeyl-5-(5-nitrofurfurylidene)-2-t;hioxothiazolidineA-one. 0 Ethylol 163 3-hydroxyethyl-5-(5-nitroturturylidene)-2-thioxo-thiazolidineA-one. 0 Cyclopropyl 154 3-cyclopropyl-5-(5-nitrofurfurylidene)-2-thioxothiazolidineA-one. 0 .l Ortho-hydroxy cyelohexyl. 203 3-ortho-hydroxyeyclohexyl-fi-(5-nitrofurfurylidene)-2:thioxothiazolidine- L-one. 1 Br H 218-220 5-[2-bromo-2-(5"-n.it;rot'nrfurylidene)ethylidene]-2-thioxo-thia zolidine-4-one. 1 H H 235 5[2-5-nitroiurfmylidene)ethy1idene]-2-thioxo-thiazolidine-4-one. 0 Carboxymethyl (ammonium salt) 230 Ammonium salt of 3earboxymethyl-5-(5-nitrofurfurylidene)-2- thioxo-thiazolidinei-one. 1 H Carboxymethyl 230 3-carboxymethyl-5-[2'-(5-nitroiuriurylidene)ethylidene1-2-thixothiazolidinel-one. O l-carboxyethyl (ammonium salt) 180 Ammonium salt of 3(1carboxyethyl)-5-(5"-nitrofurfurylidene)-2- thioxothiazolldine-4-one. 0 Carbethoxy-methyl 206 3-ethoxycarbonyhnethyl-5-(5-nitrofurfurylidene)-2-thioxothiazolidineA-one. 1 H -.-..d0 3-ethoxycarb0nylmethy1-5-[2-(5-nitrofurfurylidene)ethylidene1- Z-thioxothiazolidine--oue.

TABLE IV-Continued M.P. Example Code (deg.) Name 11175 2.28 2-u-butylamino-b-(J-nltrofurfurylidene)-thiazoline-4-one.

11176 232 2r41-butylan1ino-5[2-(5"-n.ittoiururylidene)ethylideue1-thiazolineone.

11177 295 2-eyclohexylamino-5(5-nltrofurfurylidene)-thiazoline'4-one.

11178 268 2-tlzyelfixylamlno-b[2-(5-nitrofurfurylidene)ethylidene1-thiazome he.

11378 312 2-cyelohexylamlno-5[2-bromo-2'-(5"-nitroiu.durylidene)ethylidene] thiazoline-4-one.

36 11020 254 2-benzylamino-5(5-nitroiu.rfurylidene)-thiazoline-4-one.

11021 242 2-benzylamino-5[2-(5-nitroiuriurylidene)ethylidenel-thiazoline- 4-oue.

11379 286 2-benzylamino-5[2'-bro1no-2'-(5"-nitrofi1rfurylidene)ethyl1dene]- thiazoline-4-one.

11254 271 2-para-chlorobenzylamino-5(fimitroturfdrylidene)-thiazoline-4- one.

11255 247 2-para-chlorobenzylamino-5[2-(5-nitroiurtury1idene)ethylldenelthiazoline-4-one.

11262 270 2-dimethylamino-5(5'-nitroiur1urylldene)-thiazoline-4-one.

11263 218 2-iliimethylam1n0-5[2-(5"-n1trofurfuryl1dene) ethylidene1-thiazone-4-one.

11389 256 2-dlmethylamino-5[2-bromo-2-(5-n.ltroiuz1urylidene) ethylldene]thiazolinet-one.

11273 Ethyl Ethyl 200 2-dlethylamino-5(5'-nitrofurl'urylldene)-thiazoline-4-one.

11390 1 do 228 Z-diethylamino-[2-(5"-nitroiurfurylidene) ethylideue1- thiazoline-l-one.

11391 1 Br ..do ..do-- 211 2-diethylamino-5[2-bromo-2'-(5-nitroturfurylidene) ethylidene] thiazollneA-one.

11274 0 Benzyl Methyl 224 Z-methylbenzylamino-E(5'-nltrofuriurylidene)-thiazollne-4-one.

11275 1 H .do .-do 209 2-methylbenzy1amino-5[2'-(5"-n.ltroiurfury1ldene) ethylldene1- thiazoline-a-one. I

11276 0 Phenyl .-do 240 2-methylphenylamlno-5(5-nltrofurl'urylidene)-thlazoline-4-one.

11291 1 H do 233 2-methylphenylamlno-5[2-(5"-nltroiurfurylidene)ethylidene} thlazolinel-one.

11292 0 Morphollno 278 2-(N-morpholino)-5(5'-nltrofurfurylidene)-thluzoline-4-one.

11293 1 H Morphollno 238 2-(N-morpholiuo)-5-[2(5-nltroiurfurylldene)ethylidene1- thlazoline'one.

Inst.

The following examples illustrate the preparation of the compounds.

EXAMPLE 1 Compound 10290 0.705 g. (0.05 mol) of S-nitrofurfural in solution in 5 ml. of absolute ethanol is added to 0.80 g. (0.05 mol) of 3-ethylrhodanine in solution in 5 ml. of ethanol and 1.5 g. of anhydrous potassium acetate in solution in 5 ml. of acetic acid. The reaction mixture is heated to 65 C. on a water bath for 1.5 hours; after cooling the precipitate is separated by suction filtration, washed with water, then with alcohol and ether and finally dried. The product has M.P.=150 C. Yield 1.1 g. (77% of theoretical) The derivatives which have, in the 3-position, a methyl (10289), allyl (10291), n-heptyl (10292), n-dodecyl (10293), n-hexadecyl (10381), B-hydroxyethyl (10294), cyclopropyl (10385), or 2-hydroxy-cyclohexy1 (10387) group are obtained in a similar manner by using the corresponding 3-substituted rhodanines.

EXAMPLE 2 Compound 10453 1 g. (0.004 mol) of a-bromo-p-(5-nitrofuryl-2)-acrolein is dissolved in 5 ml. of ethanol and treated with 0.53 g. (0.004 mol) of rhodanine dissolved in 5 ml. of hot ethanol and then with 1.32 g. of anhydrous potassium acetate dissolved in 6 ml. of acetic acid. After heating on a water bath at 60 C. for 4 hours, a precipitate is formed which is separated by suction filtration after cooling, washed with water and ethanol and finally recrystallised from nitromethane. The compound is a red powder which melts at 218 to 220 C. and weighs 0.7 g. (yield: 49%).

EXAMPLE 3 Compound 10452 1 g. (0.0075 mol) of rhodanine dissolved in 12 ml. of absolute ethanol and 2.2 g. of anhydrous potassium acetate dissolved in 10 ml. of pure acetic acid are added to 1.26 g. (0.0075 mol) of B-(S-nitrofuryl-Z)-acrolein dissolved in 8 ml. of absolute ethanol.

After heating on a water bath at 75 C. for 2 hours and cooling, the precipitate is separated by suction filtration, thoroughly washed with alcohol and dried. The product is obtained in the form of red needles which turn yellow when washed with a little dimethylformamide. 1.52 g. is obtained (yield 72% M.'P.=235 C.

EXAMPLE 4 Compound 10374 The following solutions are prepared: 2.82 g. (0.02 mol) of S-nitrofurfural in 20 ml. of ethanol, 3.82 g. (0.02 mol) of N-rhodanine acetic acid in 40 ml. of ethanol and 5.5 g. of anhydrous potassium acetate dissolved in 35 m1. of acetic acid. These solutions are mixed together and heated at 70 C. on a water bath for 4 hours; after removal of the major portion of the alcohol under reduced pressure, the precipitate obtained is washed with water, then with alcohol and recrystallised from nitromethane. It melts at 218 C. 4.2 g. of product is obtained (yield: 67%).

The acid is suspended in a little distilled water and neutralised to pH 7.0 with a 0.1 N ammonia solution. By lyophilisation of the solution, the ammonium salt, which melts at 230 C. with decomposition, is obtained.

Similar compounds having in the 3-position a carboxymethyl group (10483), l-carboxyethyl (10380) or carbethoxyrnethyl (10481) as well as the compound having a 2- (5"-nitrofurfurylidene) ethylidene group in the 5-position and a carbethoxymethyl group in the 3-position (10482) are obtained in analogous manner to that described above from the correspondingly substituted rhodamine derivatives.

'EXAMPLE 5 Compound 10580 3.84 g. (0.02 mol) of 3-carboxymethylrhodanine are suspended in ml. of anhydrous benzene and treated on a Water bath at 80 C. with 4.8 g. (0.04 mol) of thionyl chloride for about 30 minutes with stirring. After this time dissolution is complete and the benzene and excess of thionyl chloride are evaporated underreduced pressure; the residual oil is the crude carboxylic acid chloride.

This oil is dissolved in 50 m1. of anhydrous ether and treated, with cooling, with 2.84 g. (0.04 mol) of pyrrolidine in 20 ml. of ether. The amide which precipitates is separated by suction filtration and recrystallised from methanol. When dried, it weighs 3.2 g. (yield 62%) and melts at 212 C.

13 1.22 g. (0.005 mol) of this amide in solution in 10 ml. of acetic acid, and 0.705 g. (0.005 mol) of -nitrofurfural in solution in 5 ml. of ethanol are added to 1.5 g. of potassium acetate dissolved in 8 ml. of acetic acid. After heating for 3 hours on a water bath at 65 C., the pre- 5 cipitate is filtered off, washed with ethanol and recrystallised from a little acetic acid. The dry product weighs 1.3 g. (yield 62%) and melts at 264 C.

The derivatives having in the 3-positiou, a carbamoylmethyl (10578), a morpholino-carbamoylmethyl (10583) or isonicotinylhydrazido carboxymethyl (10586) group as well as those which have a 2-(5"-nitrofurfurylidene) ethylidene group in the 5-position and in the 3-position a pyrrolidino carbamoyl methyl group (10581), a carbamoylmethyl group (10579), a morpholino carbamoyl methyl group (10584) or an isonicotinylhydrazidocarboxymethyl group (10587) are obtained in an analogous manner from the acid chloride and ammonia, morpholine or isonicotinic acid hydrazide followed by condensation with S-nitrofurfural or fi-(S-nitrofurfuryl-Z) acrolein.

EXAMPLE 6 Compound 10683 13 g. (0.046 mol) of 2-thioxo-5[2'-(5"-nitrofurfurylidene)ethylidene]-thiazolidine-4-one are dissolved at 70 C. in 200 ml. of dimethylformamide; 0.5 ml. of piperidine followed by 8.6 ml. (0.1 mol) of 35% aqueous formaldehyde are then added thereto. After 2 minutes on a water bath at 70 C., the clear solution is kept at room temperature for 5 hours, and is then poured onto a mixture of ice and water (about 500 ml.) containing 5 ml. of 35% formaldehyde. The precipitate is filtered, washed with 1% aqueous formaldehyde and dried under reduced pressure over phosphorus pentoxide at 50 C. to constant weight. It then weighs 13 g. (90% yield), instantaneous melting point: 192 C. By progressive heating it loses one equivalent of formaldehyde at temperatures of 120 C. upwards.

The compound which has a 5-nitrofurfurylidene group in the 5-position (10682) is obtained in a similar manner from 2-thioxo-5-(5-nitrofurfurylidene)thiazolidine-4-one.

EXAMPLE 7 Compound 10540 1.28 g. (0.005 mol) of 5-(5'-nitrofurfurylidene)-2-thioxo-thiazolidine-4-one are suspended in 30 ml. of ethanol, and 0.45 ml. (slight excess) of 35% aqueous formaldehyde is added dropwise and then, while cooling on an ice bath, 0.44 g. of distilled morpholine. After 15 hours stirring at room temperature, the precipitate is filtered by suction and dried. A little more of the same product is obtained by evaporating the ethanol. There is obtained 1.7 g. (yield 96%), M.P.=156-157 C.

EXAMPLE 8 Compound 10535 A solution of 01705 g. (0.005 mol) of S-nitrofurfural in 5 m1. of ethanol is added to 1.44 g. (0.005 mol) of 3-(parasulphonamidophenyl)-rhodanine in solution in the minimum quantity of boiling acetic acid and 1.4 g. of anhydrous potassium acetate in solution in 8 ml. of acetic acid. After this reaction mixture has been heated on the boiling water bath for 4 hours, the greater part of the solvent is evaporated under reduced pressure, the precipitate is filtered by suction, washed with a little ethanol and dried. The product weighed 1.4 g. (yield 68%), M.P. (decomposition) :260 C.

The analogous derivatives having in the 3-position a para-chlorophenyl (10382), para-carboxyphenyl (10432), benzyl (10383) or para-chlorobenzyl (10384) group as well as that which has a 2'-(5"-nitrofurfurylidene)-ethyl- 'idene group in the 5-position and a para-sulphamoylphenyl group in the 3-position (10536) can be obtained by this process from the corresponding 3-(substituted phenyl) rhodanines and, in the latter case B-(S-nitrofuryl-Z) acrolein.

EXAMPLE 9 Compound 10430 A solution of 0.85 g. (0.006 mol) of S-nitrofurfural in 5 ml. of ethanol is added dropwise to 0.89 g. (0.006 mol) of 3-aminorhodanine dissolved in a mixture of 15 ml. of ethanol and 0.5 ml. of 2 N hydrochloric acid. The reaction mixture is kept on a water bath at 60 C. for

' minutes and then, after standing overnight at room temperature, the precipitate is filtered by suction and dried; it weighs 1.5 g.

The hydrazone prepared as described above (1.5 g. 0.0055 mol) is dissolved in 20 m1. of boiling acetic acid and treated with 0.85 g. (0.006 mol) of 5-nitrofurf'ural dissolved in 3 ml. of acetic acid and 2.5 g. of anhydrous potassium acetate dissolved in 15 ml. of acetic acid. After 3 hours heating on a water bath at C. and cooling, the precipitate is isolated and recrystallised from nitromethane. M.P.=256-258 C. The product weighed 1.6 g. (overall yield: 68%).

EXAMPLE 10 Compound 10743 and its potassium derivative (A) 118 g. (1.2 mol) of anhydrous potassium acetate and 234 g. (2 mol) of thiazolidine-2,4-dione are dissolved at C. in the course of 15 minutes in 750 ml. of glacial acetic acid. The reaction mixture is then left to cool to 75 C., and 126 g. (0.9 mol) of S-nitrofurfural dissolved in 250 ml. of acetic acid are then added all at once and the resulting reaction mixture heated at C. for 45 minutes. After cooling, the solution is poured onto 4 kg. of ice and left overnight at room temperature. The precipitate is filtered by suction, washed with ethanol and dried at 100 C. under reduced pressure to constant weight since the product contains 1 mol of solvent of crystallisation. The product weighed 165 g. (yield 76.5%). The M.P. is 225 C.

The same product can also be obtained quantitatively from 5-(5'-nitrofurfurylidene)-pseudothiohydantoin prepared as described in Example 27 (see below) by treating 1 g. of this product on a boiling water bath for 4 hours, with stirring, with 20 ml. of 6 N hydrochloric acid.

(B) 12 g. (0.05 mol) of 5-(5'-nitrofurfurylidene)- thiazolidine-2,4-dione are suspended in 75 ml. of pure ethanol and treated at 0 C. with a solution of 2.8 g. (0.05 mol) of pure potassium hydroxide in 50 ml. of pure ethanol. The reaction mixture is allowed to return to room temperature, and stirring continued for 2 hours. The precipitate of the potassium derivative is filtered by suction, washed with a little ethanol and dried. The product weighed 13.6 g. (yield 98%). M.P. (decomposition) 290 C.

EXAMPLE 11 Compound 10744 and its potassium salt (A) 234 g. (2 mols) of thiazolidine-2,'4-dione are added to a solution of 196 g. (2 mols) of potassium acetate in 800 ml. of acetic acid at 90 C. This temperature is maintained until the product has dissolved; 167 g. (1 mol) of B-(S-nitrofuryl-Z)-acrolein in solution in 400 ml. of acetic acid are then added. The temperature is raised to C. and held for one hour. Precipitation, which is abundant Whilst still hot, is completed upon cooling. The filtered precipitate is washed with a little acetic acid, then with Water and alcohol and finally recrystallised from about 700 ml. of dirnethylformamide. After prolonged drying in a heated oven at 100 C. under reduced pressure it melts at 260 C.; 146 g. of this product are obtained (yield 55%).

(B) 106 g. (0.4 mol) of the product prepared as described above are suspended in 500 ml. of pure ethanol and treated at 0 C. with 22.4 g. (0.4 mol) of pure potassium hydroxide in solution in 500 ml. of ethanol. After 2 hours stirring at room temperature the precipitate of the potassium salt is separated by suction filtration and dried. 120 g. of this product are obtained (yield 99%), M.P. (decomposition) =325 C.

EXAMPLE 12 Compound 11137 2.34 g. (0.02 mol) of thiazolidine-2,4-dione are dissolved in a solution of acetic acid containing 2 g. of po tassium acetate in 8 ml. of acetic acid at about 90 C. When solution is complete, 2.46 g. (0.01 mol) of 2-br omo-3-(5'-nitro-furyl-2')-acrolein in solution in 6 ml. of acetic acid are added and the reaction mixture is heated under reflux for one hour. After standing overnight at room temperature, the precipitate is separated by suction filtration, washed with a little dimethylformamide, then with ethanol and dried. The product weighed 1.2g. (yield 35%). M.P.=260 C.

EXAMPLE 13 Compound 10933 (A) 3.93 g. (0.03 mol) of 3-methyl-thiazolidine-2,4- dione in solution in 4 ml. of acetic acid are added to 4.23 g. (0.03 mol) of 5-nitrofurfural in 4 ml. of acetic acid and 5.9 g. (0.06 mol) of anhydrous potassium acetate in 12 ml. of acetic acid. The mixture is heated at 125 C. for one hour and left overnight. The precipitate thus obtained is removed by suction filtration, washed with acetic acid, water and then ethanol and finally recrystallised from dioxane. 2.33 g. (yield 31%) of a deep yellow powder melting at 208 C. are thus obtained.

(B) 1 g. (0.036 mol) of the potassium derivative of 5-(S'mitrofurfurylidene)-thiazolidine-2,4-dione prepared as described in Example is treated with 0.85 g. (0.06 mol) of methyl iodide in 10 ml. of dimethylformamide on a water bath until the reagents have gone into solution and the potassium iodide formed has completely precipitated; the dimethylformamide is then evaporated under reduced pressure, the residue treated with water and the water filtered off, and the precipitate is then recrystallised from dioxane. 0.86 g. (yield 94%) of a yellow product which melts at 211 to 212 C. is thereby obtained.

(C) 2.7 g. (0.01 mol) of 2-methylimino-3-methyl- 5(5' nitrofurfurylidene)-thiazolidine-4-one the preparation of which is described in Example 29, are treated with 50 ml. of concentrated hydrochloric acid under reflux for 7 hours. The insoluble portion is removed by suction filtration of the hot solution, Washed with water and dried. It weighs 0.8 g. and melts at 210 C.; the acid solution contains unhydrolysed starting material in the form of the hydrochloride. The product which melts at 210 C. is identical with that obtained as described under A and B.

Analogous products which carry, in the 3-position, an ethyl (11064), B-chloroethyl (11080), isopropyl (11071), allyl (10829), n-hexyl (1107 6), propynyl (11245), npentyl (11246), n-heptyl (11248), n-octyl (11250) or n-hexadecyl (11252) .group as well as those which carry a 2-(5" nitrofurfurylidene)ethylidene group in the 5- position and, in the 3-position, a methyl (10934), ethyl (11065), isopropyl (11072), allyl (10830), n-hexyl (11077), n-pentyl (11247), n-heptyl (11249), n-octyl (11251) or n-hexadecyl (11253) group are obtained by the same processes using the appropriately substituted thiazolidine 2,4 diones as starting materials with in process B the appropriate alkyl or propynyl iodide and in process C the appropriate 2-methylimino 3 alkyl- S-substituted thiazo1idine-4-ones.

EXAMPLE 14 Compound 11078 1.3 g. (0.012 mol) of ethyl chloroformate is added, with cooling on an ice bath, to a suspension of 2.78 g.

(0.01 mol) of the potassium salt prepared as described in Example 10 in 25 ml. of dimethylformamide. After one hour in the cold, the solvent is evaporated under reduced pressure at 70 C. and the residue is treated with water and recrystallised from ethanol.

The product obtained in this way melts at 123-124 C. and, after drying, weighs 1.67 g. (yield 51%).

EXAMPLE 15 Compound 10827 (A) 8.4 g. (0.06 mol) of 5-nitrofurfural are addedto a solution of 13.2 g. (0.06 mol) of 3-ethoxycarbonylmethylene thiazolidine 2,4 dione in 20 ml. of acetic acid, and 15 g. of anhydrous potassium acetate in solution in 30 ml. of acetic acid are then added at 60 C. After a few minutes heating on a water bath at 90 C., an exothermic reaction ensues and copious precipitation takes place. After prolonged cooling, the mass is treated with water and the solid present separated by suction filtration, washed with ethanol and then recrystallised from 7 ml. of dim ethylformamide, removed by suction filtration, washed with alcohol and dried under reduced pressure. 8.5 g. of product are obtained (yield 43%), M.P.=l181 C.

(B) 7 g. (0.025 mol) of the potassium salt of 5(5'- nitrofurfurylidene)-thiazolidine 2,4 dione obtained as described in Example 10 are treated with 6.12 g. (0.05 mol) of ethyl chloroacetate in 40 ml. of dimethylformamide on a boiling water bath for one hour. After removal of the dimethylformamide under reduced pressure and treatment with water, the precipitate obtained is separated by suction filtration, washed with ethanol and recrystallised from acetic acid. 7 g. of product (yield 86%) are thereby obtained. M.P.=181-182 C.

EXAMPLE 16 Compound 10828 This product is obtained by one or other of the two methods described in the preceding example, substituting fi-(5-nitrofuryl-2)-acrolein for S-nitrofurfural in part A or the potassium salt obtained as described in Example 11 for that of Example 10 in part B. The two samples obtained in this way both melt at 150 C.

EXAMPLE 17 Compound 11067 2.78 g. (0.01 mol) of the potassium salt the preparation of which is described in Example 10, 1 g.' (0.0105 mol) of monochloroacetic acid and 25 ml. of dimethylformamide are heated at 90 C. for 3 hours. After concentration of the solvent under reduced pressure and treatment with water, the residue is separated by suction filtration and recrystallised from 250 ml. of water. 1.5 g. (yield 50%) of a product which melts at 115 C. is thus obtained.

EXAMPLE 18 Compound 1 1079 The method described in the preceding example is repeated using the potassium salt obtained in Example 11, to give the desired productin 66% yield. It melts at 245 C.

EXAMPLE 19 Compound 10946 6.4 g. (0.028 mol) of 3-(3-ethoxycarbon'yl 2 propenyl)-thiazol idine 2,4 dione and 4 g. of S-nitrofurfural (0.028 mol) in solution in 6 ml. of pure acetic acid are treated with 5.5 g. (0.056 mol) of potassium acetate in solution in 11 ml. of acetic acid at C. After 2 hours at room temperature and treatment with water, the precipitate is isolated and recrystallised from acetic acid. It has M.P.=l54 C. and 2.36 g. is obtained (yield 24% 1 7 This product can also be obtained by heating the potassium salt described in Example 10 with ethyl 'y-bromocrotonate.

EXAMPLE 20 Compound 10947 Replacing S-nitrofurfural used in the preceding example by a. molar equivalent of ,3-(S-nitrofuryl-Z)-acrolein, a small yield of the desired product is obtained. The yield is almost quantitative, however, when the potassium salt obtained as described in Example 11 is heated with ethyl 'y-bromo-crotonate dissolved in dimethylformamide and proceeding-as described in Example 15(B). This product melts at 173 C.

EXAMPLE 21 Compound 10787 18 g. (0.075 mol) of (5'-nitrofurfurylidene)-thiazolidine 2,4 dione prepared as described in Example are dissolved in 75 ml. of dimethylformamide at 70 C. 0.5 ml. of piperidine are added thereto, followed by 13 ml. of 37% formaldehyde (0.15 mol). After two more minutes at 70 C. the reaction mixture is allowed to stand at room temperature for 12 hours, the solution poured on 300 ml. of a mixture of water and ice containing 10 ml. of 37% formaldehyde. The precipitate which is obtained is separated by suction filtration, washed with 1% aqueous formaldehyde, separated by suction filtration and dried in the presence of phosphorus pentoxide under reduced pressure at 100 C. The dry product weighs g. (yield 74%). When heated at progressively increasing temperatures, it loses one equivalent of formaldehyde from 120 C. upwards to yield the starting material.

EXAMPLE 22 Compound 10932 0.86 ml. (0.01 mol) of 35% formaldehyde are added at 80 C. to 1.33 g. (0.005 mol) of 5[2' (5" nitrofurfurylidene)ethylidene]-thiazolidine 2,4 dione in solution in '12 ml. of dimethylformamide. After 10 minutes at 75 C., the solution is left to stand overnight at room temperature and then treated with 3 volumes of a mixture of ice and Water; the solid formed is separated by suction filtration, washed with a little 1% aqueous formaldehyde and then dried under reduced pressure in the presence of phosphorus pentoxide.

The product obtained in this way weighs 1 g. (yield 68%). It loses formaldehyde when heated above 120 C.

EXAMPLE 23 Compound 10985 1.33 g. (0.005 mol) of 5[2'-(5"-nitrofurfurylidene) ethylidene]-thiazolidine-2,4-dione in suspension in 25 ml. of ethanol is treated with 0.45 ml. (slight excess) of formaldehyde in a 35% aqueous solution and with 0.055 mol of 15% ethanolic dimethylamine. After stirring for one hour at room temperature, the precipitate is separated by suction filtration, washed with a little ethanol and dried at 100 C. under reduced pressure. It weighs 1.2 g. and melts at 175 C.

Operating in a similar manner in dioxane or in dimethylformamide as solvents, the analogous compounds which carry, in the 3-position, a diethylaminomethyl (11136), bis(5-hydroxyethyl) aminornethyl (11003), pyrrolidinomethyl (11000), piperadinometh'yl (11004), morpholinomethyl (10889), piperazinomethyl (11075) or 4- (B-hydroxyethyl)-piperazinomethyl (11066) group as well as those which carry a 5-nitrofurfurylidene group in the 5-position and, in the 3-position, a dimethylamino methyl (11001), bis-()3 hydroxyethyl) aminomethyl (10988), pyrrolidinomethyl (1099), piperidinomethyl (10989) or morpholinomethyl (10888) group are obtained from the two starting compounds prepared as described in Examples 10 and 11 and substituting the appropriate secondary amines for dimethylamine.

EXAMPLE 24 Compound 10677 9.8 g. (0.05 mol) of the hydrochloride of 3-( 3-aminoethyl) thiazolidine 2,4dione in solution in 50' ml. of acetic anhydride are heated at C. for 3 hours; the solvent is then evaporated under reduced pressure and the acetyl derivative obtained is recrystallised from water. 6.35 g. are obtained. M.P.=116 C.

2.02 g. (0.01 mol) of this derivative and 141 g (0.01 mol) of S-nitrofurfural in solution in 10 ml. of pure ethanol are treated with 2.94 g. (0.03 mol) of potassiurn acetate in solution in 10 ml. of pure acetic acid and the whole is heated to boiling on a water bath for 4 hours. The solid which has precipitated is filtered after the reaction mixture has been allowed to cool and is recrystallised from ethanol. It is a yellow product which melts at 182 C. and weighs 0.9 g.

0.8 g. of this condensation product and 20 ml. of 4 N-hydrochloric acid are heated under reflux for 4-hours. After one night in the refrigerator, the hydrochloride is separated by suction filtration, washed with a little ice water and dried. It weighs 0.6 g. The instantaneous melting point is 205 C.

EXAMPLE 25 Compound 10741 This product is obtained from the hydrochloride of 3- (B-aminoethyl) thiaZolidine-2,4-dione and B-(S-nitrofuryl-2)acrolein by the process described in Example 24. It is soluble in cold water to the extent of 4 g. per litre. The instantaneous melting point is 220 C.

EXAMPLE 26 Compound 10831 5.56 g. (0.02 mol) of the potassium salt of 5(5'-nitrofurfurylidene)-thiazolidine-2,4-dione the preparation of which is described in Example 10, and 3.16 g. (0.025 mol) of benzyl chloride in 50 ml. of dimethylfarmamide are heated on a boiling water bath for 3 hours. After evaporation of the solvent under reduced pressure and treatment with water, the residue is removed by suction filtration and recrystallized from acetic acid. 5 g. (yield 76%) of the product which melts at 201 C. are thus obtained.

A similar compound which carries a para-nitrobenzyl group in the 3-position (10833) (from para-nitrobenzyl chloride) and those which carry a 2'-(5"-nitrofurfurylidene)ethylidene group in the 5-position and, in the 3- position, a benzyl group (10832) or a para-nitrobenzyl group (10834) are obtained in the same manner from the potassium salts described in Examples 10 and 11.

EXAMPLE 27 Compound 10746 1.67 g. (0.01 mol) of fl-(5-nitrofuryl-2)-acrolein in solution in 5 ml. of acetic acid is added to a solution of 2.44 g. (0.025 mol) of anhydrous potassium acetate and 1.16 g. (0.01 mol) of pseudothiohydantoin in 10 ml. of acetic acid at 70 C. The whole reaction mixture is kept on a water bath at this temperature for 6 hours. After prolonged cooling, the precipitate is separated by suction filtration, washed with acetic acid, then with water and lastly with ethanol and dried; the dark brown solid product thus obtained weighs 1.52 (yield 57%): it is recrystallised from dimethylformamide. In the pure state, it is an orange yellow solid which melts with decomposition at abou 300 C.

1 9 EXAMPLE 28 Compound 1 1 141 2 g. of anhydrous potassium acetate are dissolved in 8 ml. of pure acetic acid; 1.3 g. (0.01 mol) of 3-methyl- 2-imino-thiazolidine-4-one is then dissolved therein and then, after the addition of 1.41 g. (0.01 mol) of 5-nitrofurfural in solution in 2 ml. of acetic acid, this solution is heated on a water bath at 60 C. for 15 minutes. After cooling, the solid formed is removed by suction filtration, washed with a little acetic acid, then water and lastly with ethanol and dried. It can be recrystallised from dimethyl formamide. 2.45 g. (yield 97%) of the product is obtained, M.P.=259-260 C.

Compound 11142 which melts at 224-225 C. is obtained by replacing S-nitrofurfural by a molar equivalent of B-(S-nitrofurfuryl-Z)-acrolein in the above example.

EXAMPLE 29 Compound 11025 A solution of 1.41 g. (0.01 mol) of S-nitrofurfural, 1.44 g. (0.01 mol) of 2-methylimino-3-methyl-thiazolidine-4-one and 1.9 g. (0.02 mol) of anhydrous potassium acetate in 9 ml. of acetic acid is heated at 60 C. for 7 minutes. The orange yellow precipitate which forms is separated 'by suction filtration after complete cooling, and washed with water and ethanol. 2.08 g. (yield: 75%) are thereby obtained. M.P.=23'8 C.

This product may be recrystallised from dimethylformamide; the product obtained is greenish yellow. M.P.=240 C.

' Analogous compounds which have an allyl group in the 3-position and an allylimino group in the 2-position (10983) or a phenyl group in the 3-position and a phenylimino group in the 2-position (10986) as well as those which have a 2-*('5"-nitrofurfurylidene)ethylene group in the 5-position and, in the 3-position and 2- position, methyl, methyl and methylimino groups respectively (11026), or allyl and allylimino groups respectively (10984) are obtained under similar conditions but with longer time of heating starting with the correspondingly substituted thiazolidine-4-ones and in the two latter cases. B-(S-nitrofuryl-Zl-acrolein.

EXAMPLE 30 Compound 11002 1.96 g. (0.02 mol) of anhydrous potassium acetate dissolved in 4 ml. of acetic acid, and then 1.41 g. (0.01 mol) of S-nitrofurfural in solution in 3 ml. of acetic acid, are added to 1.92 g. (0.01 mol) of 2-phenylimino-thiazolidine- 4-one in solution in 3 ml. of acetic acid. The solution is heated on a water bath at 75 C. for 2 hours 30 minutes. After cooling to room temperature, the solid which separates is isolated by suction filtration, washed with water and then with alcohol, and recrystalllised from dimethylformamide. When dried the product weighs 1.3 g. (yield 41%) and melts at 315 C.

The corresponding compound carrying an ortho-chlorophenylimino group in the 2-position (11005) (using 2- (orthochlorophenylimino-thiazolidine-4-one) or that which which has a 2'-(5"-nitrofurfurylidene) ethylidene group in the 5-position and a phenylimino group in the 2- position (11019) (using ,8-(5-nitrofuryl-2) acrolein) is obtained by the same method.

EXAMPLE 31 Compound 11022 0.5 ml. of 35% formaldeyhde (slight excess) and 0.44 g. (0.005 mol) of distilled morpholine are added successively to 1.57 g. (0.005 mol) of 2-phenylimino-5(5'-nitroforfurylidene)thiazolidine-4-one (the preparation of which is described in Example 3.0) in suspension in 10 ml. of dimethylformamide.

furfurylidene) ethylidene group in the 5-position (11023) (using Z-phenylimino-S 5 '-nitrofurfurylidene ethylidene) thiazolidine-4-one and that which carries an ortho-chlorophenylimino group in the 2-position and a 5-nitrofurfurylidene group in the 5-position (1 1024) (using 2-(orthochlorophenylimino -5- (5 '-nitrofurfurylidene thiazolidine- 4-one) are obtained by applying the same technique.

EXAMPLE 32 Compound 11058 1.19 g. (0.005 mol) of 2- (para-toluene-sulphonylimino)thiazolidine-4-one, 0.705 g. (0.005 mol) of S-nitrofurfural and 0.98 g. (0.01 mol) of potassium acetateare dissolved in 6 ml. of acetic acid and heated on a water bath at 60 C. for 20 minutes. After cooling, the product is separated by suction filtration, washed with acetic acid, water and then ethanol and dried. It weighs 1.3 g. (yield: 66%) and melts at 267 C.

The corresponding compound carrying a benzenesulphonyl imino group in the 2 position (11060) (starting from 2-benzene-sulphonylimino)-thiazolidine-4-one) and the compounds which carry a 2'-(5"-nitrofurfurylidene) ethylidene group in the 5-position and, in the 2-position, a para-toluene-sulphonyl-imino group (11059 or a benzenesulphonylimino group (11061) can be obtained in the same manner starting from 8(5-nitrofuryl-2)-acrolein and the stated thiazolidine-4-ones.

EXAMPLE 33 Compound 11073 0.87 g. (0.005 mol) of the semicarbazone of thiazolidine-2,4-dione in suspension in a solution of 15 ml. of acetic acid containing -1 g. of potassium acetate is treated with 0.84 g. (0.0005 mol) of 9-(5-nitrofuryl-2)-acrolein for 30 minutes under reflux and with stirring. After the precipitate which forms has been cooled, it is filtered, washed with acetic acid and with ethanol and dried under reduced pressure. This product melts at 248 C.

EXAMPLE 34 Compound 11074 3.28 g. (0.02 mol) of Z-guanidinyl thiazolidine-4-one, 2.82 g. (0.02 mol) of S-nitrofurfural and 3.92 g. (0.04 mol) of anhydrous potassium acetate are dissolved in 60 ml. of pure acetic acid. A precipitate is formed after heating on a water 'bath at 65 C. for one hour. It is separated by suction filtration after cooling the reaction mixture, washed with ethanol and then dried. 4.18 g. (giseldcc: 71%) of the product is obtained which melts at 3 EXAMPLE 35 Compound 11 139 4 g. of anhydrous potassium acetate are dissolved in 16 ml. of acetic acid and 2.6 g. (0.02 01) of Z-methylaminothiazoline-4-one and 2.84 g. (0.02 mol) of 5-nitrofurfural are added thereto. The solution obtained is heated for 2 hours at C. on a water bath. After cooling a solid which separates is isolated by filtration and recrystallised from a small amount of dimethylform'amide; it weighs 1.2 g. and melts at 328 C.

The products which carry, in the 2-position, an ethylamino group (11062), allylamino (11171), n-propylamino (11143), isopropylamino (11173), n-butylamino 1117s or cyclohexylamino 11177 are obtained by replacing the 2-methylamino thiadolidine-4-one by the appropriate 2-alkylamino, 2-cycloalkylamino or 2-alkenylamino derivative, and compounds 11140 (2-methylamino), 11063 (Z-ethylamino), 11172 (2-allylamino), 14144 (2-n-propyla mino), 11174 (2-isopropylamino), 11176 (Z-n-butylamino) and 11178 (2-cyclohexylamino) are obtained *by replacing -nitrofurfural by 5-(5-nitrofuryl)-2)acrolein in the above described process. Starting from 2-methylamino-, 2-ethylamino-, 2-isopropylaminoand 2-cyclohexylamino-thiazoline-4-ones and a-bromo-fl- (5-nitrofury1-2)-acrolein, the compounds 11376, 11138, 11377 and 11378, respectively, are obtained.

EXAMPLE 36 Compound 1 1020 1.1 g. (0.0078 mol) of 5-nitrofurfural, 1.6 g. (0.0078 mol) of 2-benzylamino-thiazoline-4-one and 1.5 g. of anhydrous potassium acetate are successively dissolved by heating in '5 ml. of acetic acid 'and heating is continued at 65 C. for 2 hours on a water bath. After prolonged heating and treatment with water, the precipitate is removed by suction filtrate, washed with ethanol and recrystallised from a small amount of dimethylformamide. It weighs 1 g. (yield: 39%) and melts at 254 C.

By following the above described process, the corresponding compounds which have a para-chlorobenzylamino (11254), dimethylamino (11262), diethylamino (11273, 11390 and 11391), methylbenzylamino ('11274), methylphenylamino (11276) or morpholino (11292) group in the 2-position may be obtained using as starting materials the correspondingly Z-substituted thiazoline-4- ones. Those which have a 2-(5"-nitrofurfurylidene)ethylidene group in the 5-position and, in the 2-position, a benzylamino (11021), para-chloro benzylamino (11255), diethylamino (1-1263 and 11389), methylbenzylamino (11275), methylphenylamino (11291) or morpholino (11293) group are similarly obtained from fl-(S-nitrofuryl-Z) acrolein and the corresponding Z-substituted amino-thiazoline-4-ones. The homologous compound (11379) is obtained from oz-bromo, 8-(5-nitrofuryl-2)- acrolein and 2-benzylamino-thiazoline-4-one.

EXAMPLE 37 Compound 12004 (a) 15.5 g. (0.1 mol) of thiazolidine-2,4-dione potassium salt suspended in 60 ml. of anhydrous dirnethylformamide are treated with 18.8 g. (0.1 mol) of 1,2- dibromoethane on a boiling water-bath with stirring for 4 hours. Potassium bromide is separated by suction filtration, the solvent driven off in vacuo, the residual oil treated with water, extracted with chloroform, dried over magnesium sulphate then distilled; 16.40 g. of 3-{3-bromoethyl-thiazolidine-Z,4-dione are obtained (yield 73%) BP =125--130 C.; n =1.5778.

(b) 6.46 g. (0.066 mol) of anhydrous potassium acetate, 7.4 g. (0.033 mol) of 3-B-bromoethy1-thiazolidine- 2,4-dione and 4.65 g. (0.033 mole) of S-nitrofurfural are dissolved successively in 30 m1. of anhydrous acetic acid on a water-bath at 90 C.

Heating is continued at this temperature for 3 hrs. 30 min. After prolonged cooling, the precipitate is separated by suction filtration in vacuo, washed with a little ethanol and dried in an oven at 60 C. By adding a mixture of equal parts of ethanol and ice to mother liquors, a second crop is obtained, isolated in the same manner as the first one and added thereto. The product is purified by recrystallisation from benzene in the presence of animal charcoal. The yellow product thus obtained melts at 134 C. and after drying weighs 6 g. (yield 52% EXAMPLE 38 2.78 g. (0.01 mol) of 5-(5'-nitrofurfurylidene)-thiazolidine-2,4-dione potassium salt prepared according to Example 10 and 3.76 g. (0.02 mol) of 1,2-dibromethane in 25 m1. of dimethylformamide are kept on a waterbath with stirring at C. for 3 hours. After removing the solvent in vacuo, the residue is treated with Water; the precipitate is separated by suction filtration, dried and recrystallized, from benzene in the presence of animal charcoal. The product thus obtained weighs 1.65 g. (yield 48%) and melts at 132-133 C. It is the same as the product obtained according to Example 37.

EXAMPLE 39 Compound 12003 (a) 3 -chloropropyl-thiazolidine-2,4-dione is obtained in the same conditions as the fi-bromoethyl homologue according to Example 37 by treating thiazolidine-2,4 dione potassium salt with 1-bromo-3-chloropropane in stoichiometric amount. The yield of distilled product is 88%. BP 0.05 mm.=134 C.

(b) By substituting 3-' -chloropropyl-thiazolidine-2,4- dione for 3-;3-bromoethyl-thiazolidine-2,4-dione in Example 37 in an equivalent amount (6.40 g.; 0.033 mol) and using the same reaction conditions, compound 12003 melting at C. can be obtained.

The same compound may also be obtained as in Example 38 by treating 5-(5-nitrofurfurylidene)-thiazolidine-2,4-dione with an equivalent of 1-bromo-3-chloropropane. The product thus obtained melts at 158 C. and is the same as the product obtained by condensing 5- nitrofurfural with thiazolidine-2,4-dione previously substituted in the 3-position.

The compounds prepared according to the examples were subjected to biological tests, the results of which will now be described.

(1) Antibacterial action in vitro: The technique employed in the study of the products is the inhibition of growth by serial dilutions in Petri dishes containing agar medium.

Into each Petri dish are introduced 2 ml. of the intermediate range of products and 18 ml. of molten agar, and the dish is heated to 50 C. When the agar has set, the surface is dried at 37 C. The cultures, of 21 strains of bacteria and of 44 strains cultivated for 18 hours in ordinary broth are diluted to A in distilled water and used for inoculation. The dishes after turning upside down are warmed to 37 C. in a heating cupboard for 24 hours and the readings are then taken.

(2) Action against Moch Bacillus in vitro:

The liquid medium used for the tests on the serum was Youmans medium which was placed in tubes measuring 22 mm., in portions of 5 ml. per tube. The substance to be tested is distributed among the tubes in such a Way that the concentrations range from 10 mcg. to 0.05 mg. per ml. The medium is inoculated with the Mycobacterium tuberculosis H 37 R V strain from a culture 7 to 9 days old on Dubos medium, carrying out two inoculations (0.01 mg. of bacilli and 0.1 mg. per 5 ml. of medium).

The culture is titrated by nephelometry, comparing the culture in Dubos medium with that of a range covering a series of suspensions of ECG.

Readings are taken on the sixth, twelfth and eighteenth day. The bacteriostatic concentration is taken as that concentration which prevents the appearance of a culture which is visible to the naked eye on the sixth day after inoculation with 0.01 mg.

The reading taken on the eighteenth day, especially that with the 0.1 mg. inoculation, enables one to have an approximate estimate of the number of resistant bacilli present in a normal strain.

(3) Antifungal activity in vitro: Study of the antifungal properties of the products of the present invention was carried out by inoculation with difierent fungi and yeasts of a Sabouraud medium containing dilutions of the products ranging from 0.5 to 30 mcg. per ml. of solidified medium. The reading was taken after the specimen had been in a heating cupboard at 27 C. for 5 days.

23 (4) Antiparasiticidal action in vivo: (8.) Hymenolepis nana var. Fraterna: Eight weeks old mice were given 100 eggs by the oral route. Four weeks later, the animals were treated with the test material for three days. Twenty-four hours after the last'treatrnent, 5

the mice were sacrificed and the percentage of depara sitisation noted.

(b) Syphacia obvelate: Four weeks old mice were left in contact with strongly parasitised mice for eight days. The animals were then treated with the test material by the oral route for 4 days.

Forty-eight hours after the last treatment, the mice were sacrificed and the percentage of deparasitisation noted.

(c) Nippostrangylus muris: Three to four weeks old rats were given 200 larvae of Nippostrongylus subcutaneously. n the tenth, eleventh and twelfth day after inoculation, the animals were treated orally.

On the fourteenth day after inoculation, they were sacrificed and the number of adult parasites counted. The following formula gives the percentage of deparasitisation:

N n N N and n being the mean number of worms found in the control animals and the treated animals, respectively.

Urinary excretion of the products in the rat: The method of Hartley, modified by Chabbert (1957) was used for biological dosage. It consists in horizontal diffusion from discs impregnated with urine on an agar medium inoculated with the test bacteria.

The percentage of excretion is calculated from the reading of the zone of inhibition of growth of the bacteria in relation to a range of the product. Graphs are 24 drawn up in which the concentrations are entered as abscissae and the diameters of the range as ordinates. The activity of the sample to be studied is read directly from the graph obtained in this way. The specimens of urine were taken one, two, four, six, eight and twentyfour hours after a single dose of the product.

(6) Intestinal antiseptic action in the healthy rat: The excrements of control animals and of animals treated with the test compounds are mixed with distilled water to a known volume by stirring.

From this solution, which is considered the parent solution, dilutions of between 10"" and 10- are prepared. 1 millilitre of each dilution is filtered through a millipore filter of 0.45 and the filter is placed on an agar medium. After 24 hours at 37 C., the number of colonies per filter are counted. The excrements are taken for analysis twentyfour hours before the first treatment, twentyfour hours after treatment, at the third treatment, at the seventh treatment and three days after the treatment.

The number of colonies in each dilution after administration of the product to be examined is expressed as a percentage of the corresponding number before administration.

(7) Intestinal antiseptic action in the rat infected with Salmonella typhimurium: The procedure is the same as that described above but the animals are orally 1 ml. of an 18 hour culture of Salmonella typhimurium mixed with 1 ml. of egg yoke.

The filters are placed on a special Salmonella shigella (S.s.) medium. Dilutions of the excrements are 10 5 10* and 10- (8) Determination of the acute toxicity in mice by the intraperitoneal or oral route: The results obtained with the biological tests described above are shown in the following tables, numbered 1 to 8.

TABLE 1.ANTIBACTERIAL ACTION IN VIIRO [Minimum inhibiting concentrations in mcg./m1.]

Staphyloccus londonii $0. 05 $0. 05 1. 56 0. 4 0 8 0. 4 0. 8 0. 8 Staphylococcus aureus 521 IP- 0. 2 0 1. 56 1. 56 12. 5 Streptococcus pyoaenes gr. A 561 I 3. 12 25 3. 12 Streptococcus agalactlae gr. B 118 3.12 25 3. 12 50 Streptococcus faecalis gr. D A 23- 3. 12 12. 5 6. 25 50 Sarcina lutea ATGC 9 341-.-.- 1. 56 12.5 12. 5 50 Bacillus cereus ATCC 9 634 1. 56 1. 56 1. 56 1. 56 Bacillus subtilis ATCC 6 633- 0. 4 0. 8 0. 8 6. 25 Listeria monocytogenes 5 734-. 1. 56 12. 5 6. 25 25 Corynebacterium, abb0tt 12. 5 12. 5 50 Past. septica type C 5621 0.1 0.1 1. 56 1. 56 25 Post. pseudo tuberculosis C 114. 0.4 3. 12 25 12.5 50 Esch. coli A 223 11? 3. 12 6. 25 25 25 50 Each. 6011 F 260 0. 1 1. 56 50 50 50 Each. coll Monod D 198 1. 56 0. 8 25 25 50 K lebsiella pneumoniae ATCC 10 031-. 3. 12 6. 25 1. 56 3. 12 0. 8 Klebsiella mistral E 156; 50 50 50 Salmonella typhimurlum A 222 IP. 6. 25 6. 25 50 50 50 Salmonella para B E 118 3. 12 3.12 50 50 50 Shr'gclla dysenteriae 5 728 IP. 1. 56 3. 12 25 12. 5 50 Pseudomonas aeruginosa A 22 IP 50 50 50 50 50 Staphylococcus londonil 0. 2 0. 1 0. 1 Staphylococcus aureus 521 IP 0.4 0.4 0. 8 Streptococcus pyoaenes gr. A561 IP. 0. 2 12. 5 12. 5 Streptococcus agalactiae gr. B 55 118 IP. 0. 2 12.5 12.5 Streptococcus faecalis gr. DA 23 0. 8 6. 25 12. 5 Sarcz'na lutea ATCC 9 341..- 0. 8 50 50 Bacillus cereus ATTO 9 634. 0. 4 0. 4 0.4 Bacillus subtilis A'ICO 6 633.- 0.2 0. 1 0. 1 0. 8 3. 12 12. 5 0. 4 50 50 50 0. 8 1. 56 50 12. 5 3. 12 50 1. 56 12. 5 50 3. 12 12. 5 50 3. 12 12. 5 50 12. 5 12. 5 Klebsiella mistral E 156 Salmonella typhlmun'um A 222 IP 50 50 50 50 50 12.5 50 Salmonella para BE 118. 50 50 50 12. 5 12. 5 Shlgella dysenteriae 5 728 IP 12. 5 50 12.5 25 50 25 12. 5 Pseudomonas aeruqinosa A 2211 50 50 50 50 50 TABLE 1Continued Staphylococcus londouiz' 0. 4 0. 2 0. 1 $0. 50. 05 Staphylococcus aureus 521 IP 0. 8 0. 4 0. 2 Streptococcus pyooenes gr. A 561 IP 6. 25 2. 12 1. 56 1. 56 Streptococcus agalactiae gr. B 55 118 IP 6. 25 3.12 1. 56 1. 56 Streptococcus faecalis gt. DA 23 12. 5 0.2 1. 56 1. 56 Sarcina lutea ATCC 9 341 50 3. 2 12. 5 6. 25 Bacillus cereus ATCC 9 634 6. 25 1. 56 0.2 $0. 05 Bacillus subtilis ATCG 6 633 0. 8 .1 1. 56 50. 05 Listeria 5 734 3.12 1.56 0.8 0.4 C'orynebacteriurn abbott 25 25 Past. septica type C 5621 3.12 1. 56 0. 1 0. 05 0. 4 Past. pseudo tuberculosis C 114 25 1.56 3. 12 1. 56 Esch. coli A 223 IP 50 6. 25 3. 12 0. 8 0. 8 Esch. coli F 260.v 12. 5 3. 12 1. 56 Esch. coli monod 6. 25 3. 12 0. 8 0. 8 K lebsiella pneumoniae ATCC 10 03 3.12 12. 5 1. 3.12 6. 25 1. 56 Klebsiella mistral E 156 50 3. 12 6. 25 Salmonella typhirnuriu'm A 222 IP- 50 25 1. 56 50 50 6. 25 1. 56 3.12 Salmonella para BE 118 12. 5 1. 56 3.12 6. 25 3.12 Shigella dysenteriae 50 6. 25 1. 56 50 50 1. 56 0. 4 3. 12 Pseudomonas aeruainosa A 22 IP 50 50 50 50 50 Slaphyloccocus londonii 0. 1 50. 05 0. 1 $0. 05 0. 4 50. 05 0. 2 50. 05 Staphylococcus aureus 521 IP 0. 4 0.2 0. 8 0. 2 0. 8 0.2 0. 8 0.2 Streptococcus pyogenes gr. A 561 IP 3.12 1. 56 3. 12 1. 56 6. 25 3.12 6. 25 3. 12 Streptococcus agalactiae gr. B 55 118 IP. 3.12 1. 56 3. 12 1. 56 6. 25 1. 56 6. 25 3. 12 Streptococcus faecalis gr. DA 23 3. 12 1. 56 3. 12 1. 56 12. 5 1. 56 12. 5 1. 56 Sarcina lutea ATCC 9 341 50 12. 5 25 25 50 50 50 50 Bacillus cereus ATCC 9 634 0. 4 50. 05 0.2 0. 2 0. 8 0.1 3. 12 0. 1 Bacillus subtilis A'ITCC 633 0.1 50. 05 0. 4 50. 05 O. 4 $0. 05 0. 2 50. 05 Listeria nonocytogenes 5 734 0. 8 0. 4 1. 56 0. 4 3. 12 0. 4 1. 56 0. 4 Corynebacterium abbott 50 3. 12 50 50 50 50 50 50 Past. Septica type C 5621 1. 56 0. 2 0. 4 0. 8 3. 12 0. 8 1. 56 1. 56 Past. pseudo tuberculosis C114 1. 56 3. 12 1. 56 3. 12 6. 25 3. 12 3.12 0. 8 Esch. coli A 223 IP 1. 56 1. 56 1. 56 0. 4 6. 25 0. 8 3. 12 1. 56 Each. coli F 260 1. 56 1. 56 3. 12 O. 8 6. 25 0. 8 6. 25 0. 8 Esch. coli Monod D 198 1. 56 0. 8 3. 12 0. 8 6. 25 0. 8 6. 25 1. 56 Klcbsiella pneumoniae ATCC.10 031- 3. 12 1. 56 3. 12 1. 56 12. 5 0. 8 6. 25 1. 56 K'lebsiella Mistral E 156 25 6. 25 25 12. 5 50 6. 25 50 12, 5 Salmonella typhimurium A 222 IF. 3. 12 3.12 6. 25 3. 12 12. 5 1. 56 12. 5 3. 12 Salmonella para BE 118 3. 12 1. 56 3. 12 1. 56 6. 25 1. 56 6. 25 3. 12 Shigella dysenteriae 5 728 IP 1. 56 0. 8 1. 56 1.56 3. 12 1. 56 3. 12 3. 12 Pseudomonas aeruainosa A 22 I1 50 50 50 50 50 50 50 50 Staphylococcus Zondonii 0. 1 0. 05 $0. 05 $0. 05 $0. 05 0. 4 0. 1 0 Staphylococcus aureus 521 IP 0.2 0.2 0. 2 0. 1 0. 4 0. 8 0. 4 0. 8 Streptococcus pyogenes gr. A 561 IP 1. 56 0.4 1. 56 O. 8 6. 25 50 0. 8 25 Streptococcus agalactiae gr. B 55 118 IP 1. 56 0. 8 1. 56 0. 8 6. 25 50 0. 8 25 Streptococcus faecalis gr. DA 23 1. 56 0. 8 1. 56 0. 8 12.5 50 3.12 25 Sarcina lutea ATCC 9 341 6. 25 6. 25 25 12. 5 3.12 12. 5 Bacillus cereus ATCC 9 634 1. 56 0. 4 0. 1 50. 05 0. 8 3. 12 0. 1 1. 56 Bacillus subtilis ATCC 6 633 0. 1 S0. 05 go. 05 50. 05 O. 8 0. 2 50. 05 0. 4 Listeria monocytogertes 5 734 0. 4 0. 4 0. 8 0. 4 1. 56 12. 5 0. 8 1. 56 Coryrtebacterium abbott 1 12. 5 6. 25 12. 5 12. 5 6. 25 12. 5 12. 5 Past septica type C 5621 $0. 06 $0. 05 0.4 0. 4 0.1 0. 8 0. 8 1. 56 Past pseudo tuberculosis C 114. 6. 25 3.12 1. 56 12. 5 0. 8 0. 8 0. 2 1. 56 Esch. coli A 223 IP 3.12 1. 56 0. 8 O. 8 0. 8 3.12 0. 4 12.5 Esch Coli F 260- 1. 56 1. 56 0. 8 3. 12 1. 56 3.12 3. 12 12. 5 Esch. coli mond D 1. 56 0. 8 0. 8 0. 8 1. 56 8.12 0. 8 6. 25 Klebsiella pneumoniae ATCC 10 03 3. 12 1. 56 1. 56 1. 56 0. 2 0. 2 0. 05 0. 1 Klebsiella mistral E 156 M 1. 56 1. 56 6. 25 12. 5 0. 8 25 Salmonella typimurium A 222 IP 3.12 3. 12 1. 56 3.12 1. 56 3. 12 3.12 25 Salmonella para BE 118 1. 56 1. 56 1. 56 3.12 1. 56 6. 25 1. 56 25 Shigella dysenteriae 5 728 IP 1. 56 0. 4 3.12 3.12 0. 4 0. 8 0. 2 3. 12 Pseudomonas aeruginosa A 22 IP 50 50 50 50 50 50 50 Staphylococcus Zondonii $0. 05 0. 8 1. 56 1. 56 0. 10 50. ()5 0. 2 0. 2 Staphylococcus aureus 521 IP 0. 4 3. 12 6. 25 50 3. 12 1. 56 0. 2 0. 8 Streptococcus pyogenes gr. A 561 IP 0. 8 50 50 50 50 50 50 50 Streptococcus aqolactiae gr. B 118 I 0. 8 50 50 50 50 50 50 50 Streptococcus faecalis gr. DA 23 0. 8 50 12. 5 50 50 25 6. 25 50 Sarcina lutea A'ICO 9 341 50 25 25 50 50 50 12. 5 50 Bacillus cereus ATCO 9 634 0. 1 3.12 0. 8 50 50 0. 8 0. 8 3. 12 Bacillus subtilis AICC 6 633-- 0. 1 3. 12 0. 8 1. 56 0. 4 0. 8 0.2 1. 56 Listeria monocytogenes 5 734 0.4 6. 25 3. 12 50 25 0. 8 0. 8 25 Coryrtebacterium abbott 50 50 50 50 50 50 50 50 Past. Septica type C 5621 0. 8 25 25 5O 50 50 3. 12 50 Past. pseudo tuberculosis C 114. 0. 4 50 50 50 50 50 12. 5 50 Esch. Coli A 223 I 1. 56 50 50 50 50 50 25 50 Esch. Col F 260 50 50 50 50 50 50 50 50 Esch. Coli Monod D 198 0. 8 50 50 50 50 50 50 50 Klebsiclla pneumoniae ATCC 10 031. 50 12. 5 50 50 50 50 50 50 Klebsiello mistral E 156 50 50 50 50 50 50 50 50 Salmonella typimuriurn A 222 I 50 50 50 50 50 50 50 50 Salmonella para BE 118. 50 50 50 50 50 50 50 50 Shigella dysenteriae 5 728 IP 1. 56 50 50 25 50 50 12. 5 50 Pseudo'monas aeruai'aosa A 22 IP 50 50 50 50 50 50 50 50 Stgphyloccus Condonii 0. 05 0. 1 3. 12 0. 2 6. 25 0. 4 0. 1 0. 1 Staphylococcus aureus 521 IP 0. 8 0.2 3.12 0.2 6. 25 0. 4 0. 2 0. 1 Streptococcus pyogeues gr. A 561 IP 50 50 1. 56 12. 5 1. 56 12. 5 50 0. 8 Streptococcus agalactiae gr. B 55 118 IP 50 50 1. 56 12. 5 1. 56 12. 5 50 0. 8 Streptococcus paecalis gr. DA 23 25 50 25 12. 5 25 25 50 0. 8 Sarcina lutea ATCC 9 341 50 50 50 25 50 50 50 50 Bacillus cereus A'TCC 9 634..- 0. 4 0. 8 25 0. 8 25 1. 56 0. 4 0.2 Bacillus subtilis ATCC 6 633- 0. 4 0. 4 50 3.12 25 1. 56 0. 2 0. 1 Listeria 5 734-.- 3. 12 1. 56 25 0. 8 25 3. 12 3. 12 25 Corynebacterium abbott 50 50 50 12. 5 50 6. 25 50 50 Past. septica type C 5621. 50 3. 12 3. 12 0. 8 6. 25 0.4 1. 56 0. 1

TABLE 1'Continued Staphylococcus loudoniL. 0. 4 0. 2 go. 05 Staphylococcus aureus 521 I 0. 8 0.4 0. 2 Streptoccocus pyogeues gr. A 561 25 1. 56 1. 56 Streptococas algalactiae gr. B 55 118 IP-. 25 1. 56 0. 4 Streptococcus faecalis gr. DA 23 25 1. 56 1. 56 Sarciua lutea ATOC 9 341.. 25 5O 6. 25 Bacillus cereus ATCC 9634. 1. 56 0.4 0.2 Bacillus subtillis ATCC 6 633- 3.12 0. 1 go. 05 Listeria mouocytoycues 5 734- 6. 25 0.8 0.8 Coryuebacterium abbott--.. 12. 5 50 12.5 Past. septica Type C 5 621 1. 56 0.4 0. 1 Past. pseudo tuberculosis C 114-. 50 50 0.8 Esch. coli A 223 IP.-.. 50 50 6. 25 Esch. coli F 260 50 50 12.5 Esch. coli Monod D 198 50 50 6. 25 Klebsiella pueumouiae ATCC 10 031.. 50 6. 25 1 56 Klebsiella mistral E 156 50 50 50 Salmonella typhimurium A 222 1? 50 50 12.5 Salmonella para BE 118 50 50 12.5 Shiyella dysc'ntcriae 5 728 IP 25 50 3. 12 Pseudomouas aeruoiuosa A 22 IP 50 50 50 Staphylococcus loadouii.. 0. 2 0. 2 go. 05 Staphylococcus 209 IP.. 3. 12 50 0. 0. Staphylococcus aureus gervais D 319 50 50 0. 0. Staphylococcus aureus E 120 50 50 0. 0. Staphylococcus albus P C 1 200 0 0 Streptococcus pyogeries gr. A 561 IP Streptococcus pyoaeues gr. A D 313 3, 12 6. 25 Streptococcus aqalactiae gr. B.

12. 12. 5 Streptococcus pyoyenes gr. 0 12.5 12.5 Streptococcus faecalis var. Zymogenes- 50 50 Streptococcus faecalis gr. D 5 434 IP... 50 50 Streptococcus faecalis gr. D F 262-.. 50 50 Sarciua lurea ATCC 9341 50 50 Listeria monocytogenes 5 844 IP 50 50 Listeria monocytouenes Ramissa type IV.-.. Listeria monocytoyeues Joubert E 102 Eri/sipelothriz insidiosa E 159 Erysipelothriz iusidiosa A 27-- Bacillus cereus ATCO 702. Bacillus cereus AICC 9 634.. 6. 3. 12 Bacillus subtilis Leugar A 156- 1. 56 3. 12 Bacillus subtilis Searle 12.5 25

Pasteurella septica type B 55 129 Pastuerella septica type C 5 621 Pasteurella pseudotuberculosis C 114 Pasteurella J oubert E 99. Escherichia coli A 223 IP.. Escherichia coli Monod D 1 Escherichia coli 111 B4 A Escherichia coli 26 B6 A... Escherichia coli 55 B5 A. Escherichia coli ATCC 10 536.

Klebsiella pneumoniae E 155 Aerobacter aeroyenes E 115. Citrobacter Vi Salmonella typhi O 901 Salmonella para A 220 IP Salmonella para B A 214 IP Salmonella pullorum gallinarim 465 Shigella dysenteriac 5 728 IP- Shegella soimei D 374.... Morazella lwojfi D Pseudomonas aeruyinisa B 74..

Bordetella bronchiseptica.

Staphylococcus londouii. 0. 2 0. 8 Staphylococcus 209 IP. 0. 2 6. 25 0. 8 Staphylococcus aureus yervais D 319. 0.4 3.12 0. 8 Staphylococcus aureus E 120 0.4 1. 56 0.8 Staphylococcus albus P C 1 200.. 0.1 1. 56 0.4 Streptococcus pyogc'aes or. A 561 IP. 1. 56 1. 56 0. 2 Streptococcus pyogenes g'r. A D 313 1. 56 1. 56 0.8 0. Streptococcus agalactiae gr. 13..-- 3. 12 25 0. 2 Streptococcus pyogeues gr. 0 3. 12 0.8 0.2 Streptococcus faecalis var. Zymogenes. 1. 56 50 0. 8 0.8 Streptococcus jaecalis gr. D 5 434 IP 1. 66 25 0. 4 0. 8 Streptococcus faecalis gr. D F 262 1. 56 50 25 0. 8 Sarciua lutea AICC 9341-.. 3. 12 6. 25 56 12.5 Listeria monocytoyenes 5 844 1. 56 6. 25 0.8 0. 4 Listeria mouocytoaeaes Ramisse e IV. 6. 25 0. 8 0.4 Listeria monocytogeues .loubert E 102 6. 25 0.8 0.4 Erysipelothrix insidiosa E 159 0. 4 0. 4 go. 05 Erysipelothriz iusidiosa A 27.. 0. 2 0. 4 $0. 05 Bacillus cereus ATCC 10 702.. 0. 4 3. 12 0. 4 1.05 Bacillus cereus ATCC 9 634. 0.8 3. 12 0.4 0.2 Bacillus subtilis Leugar A 156. 0. 05 6. 25 0. 4 $0. 05 Bacillus subtilis Searle 0.8 6. 25 go. 05 Pasteurella scptica type B 129 1. 56 0. 2 0.05 .50. 05 Pasteurella septica type C 5 621-.. 0. 2 0.8 0.8 0. 1 Pasteurella pseudotuberculosis C 114. 3. 12 3.12 1. 56 1. 56 Pasteurella Joubert E 99 1. 56 3.12 1. 56 3. 12 Escherichia coli A 223 IP-- 6. 25 3. 12 1. 56 0. 8 Escherichia coli Monod D 198. 0.8 6. 25 1. 56 0. 8 Escherichia coli 111 B4 A-- 0.8 6. 25 6. 25 1. 56 Escherichia coli 26 B6 A- 1. 56 6. 25 1. 56 3.12 Escherichia coli 55 B5 A-.. 1. 56 6. 25 1. 56 1. 66 Escherichia coli ATOC 10 536- 1. 56 6. 25 3.12 0.8 Klebsiella pneumoniae E 155. 6. 25 50 50 12.5 Aerobactcr aeroyenes E 115 6. 25 6. 25 1. 56 1. 56 Citrobacter Vi 1. 56 12.5 3. 12 3. 12 Salmonella typhi O 901 3. 12 6. 25 3. 12 6. 25

TABLE 1 Cntinued Salmonella para A A 220 IP. 3. 12 25 12.5 Salmonella para B A 214 IP- 3. 12 25 3. 12 Salmonella pullorum oallinarum 6. 25 6. 25 3. 12 Shiaella dysenteriae 728 IP. 0. 4 3. 12 0. 8 Shiqella sonnei D 374 1. 56 6. 25 1. 56 Moraxella lwoffl D 50 50 50 Pseudomonas aeruginisa B 50 50 50 Bordetella bronchiaeptica 25 25 Staphylococcus londonii g0. 05 go. 05 0. 1 0. 2 $0. 05 Staphylococcus 209 IP 0.8 0.8 0. 8 0. 4 0. 4 Staphylococcus aurcus gervais D 3 9.. 0. 4 0. 4 0. 1 0.4 0.4 Staphylococcus aureus E 120. 0. 4 0. 2 0.2 0. 4 0. 2 Staphylococcus albus P O 1 200... 3.12 0. 1 0. 1 0. 2 Streptococcus pyogenes gr. A 561 IP-. 3. 12 1. 56 1. 56 3. 12 Streptocfccus pyoyenes gr. A D 813. 1. 56 0. 2 0.4 1. 56 Streptococcus agalactiae gr. B-... 3. 12 1. 56 1. 56 3. 12 1. 56 Streptococcus pyogenes gr. 0 3. 12 0. 4 0.4 1. 56 8 Streptococcus faecalis var. Zymogenes.. 3. 12 1. 56 1. 56 3 12 1. 56 Streptococcus faecalis gr. D 5 434 IP.- 3. 12 1. 56 1. 56 3 12 1. 56 Streptococcus faecalis gr. D F 262-- 3. 12 1. 56 1. 56 3 12 1. 56 Sarcina lutea ATCC 9341 3. 12 12. 5 50 50 25 Listeria monocytogenes 5 844 IP 1. 56 0.8 1. 56 3. 12 0.8 Listeria monocytogenes Ramisse type IV- 0.8 1. 56 3. 12 0. 8 Listeria monocytooenes Joubert E 102. 0. 8 1. 56 3. 12 0. 8 Erysipelothrix insidiosa E 159 0. 1 50. 05 0. 1 s0. 05 Erysipelothriz insidiosa A 27..- 0. 1 1. 56 12. 5 3. 12 Bacillus cereus ATGC 10 702. 0. 2 =0. 05 0. 1 0. 2 0. 1 Bacillus cereus ATCC 9 634. 0. 8 go. 05 0. 4 0. 4 0. 1 Bacillus subtilis Leugar A 156. 0. 4 1. 56 50. 05 0. 4 0. 1 Bacillus subtilis Searle 0. 1 1. 56 go. 05 0. 2 0. 1 Pasteurella septica type B 55 129- 1. 56 go. 05 0.2 0.4 0. 1 Pasteurella septica type C 5 621--.- 50. 05 go. 05 0.4 0.8 0. 8 Pasteurella pseudotuberculosis C 114. 1. 56 3. 12 3. 12 3. 12 3. 12 Pasteurella Joubert E 99 6. 6. 25 6. 25 6. 25 3. 12 Escherichia coli A 223 IP 3. 12 0. 4 0.8 1. 56 0. 8 Escherichia coli Monod D 198-... 3. 12 0.8 0. 8 1. 56 0.8 Escherichia coli 111 B4 A 3. 12 l. 56 0. 8 3.12 1. 56 Escherichia coli 26 B6 A 3. 12 0.8 1. 56 1. 56 1. 56 Escherichia coli 55 B5 A 3. 12 1. 56 1. 56 3. 12 3. 12 Escherichia coli ATOC 10 536 3.12 0. 8 0. 8 1. 56 0. 8 Klebsiella p E 155 .1. 56 6. 25 12. 5 25 12. 5 Aerobacter aerogenes E 115 12. 5 0. 8 1. 56 3.12 0. 8 Citrobacter Vi 3. 12 1. 56 3. 12 6. 25 3. 12 Salmonella typhi O 901 1. 56 6. 25 6. 25 6. 25 12.5 Salmonella para A A 220 IP 3. 12 12. 5 25 6. 25 25 Salmonella para B A 214 IP- 3. 12 6. 25 3. 12 12. 5 6. 25 Salmonella pullorum gallinarum 465.. 3. 12 6. 25 6. 25 3.12 12. 5 Shigella dysenterias 5 728 IP 1. 56 0. 8 3.12 1. 56 3. 12 Shigella sonnei D 374 1. 56 1. 56 1. 56 1. 56 3. 12 Morarella lwofli D 50 50 50 50 P ac: B 74 50 50 50 50 50 Bordetella br ptica-... 1. 56 50 25 25 5 1 1 0.05 $0.05 0.1 $0.05 $0.05 Staphylococcus 209 IP 0.4 0.4 0.4 0.2 0.4 Staphylococcus aureus aervais D 319 0. 4 0. 4 0. 4 0. 2 0. 4 Staphylococcus aureus E 120 0. 2 0. 2 0.4 0. 2 0. 2 Staphylococcus albus P C 1 200. 0. 1 0. 1 1. 56 0.8 0. 2 Streptococcus pyoyenes gr. A 561 IP. 1. 56 1. 56 1. 56 0. 4 0. 2 Streptococcus pyoaenes gr. A D 313 0. 4 0. 4 0. 4 0.1 0. 2 Streptococcus aoalactiae gr. B 3. 12 1. 56 1. 56 0. 8 1. 56 Streptococcus yoqenes gr. 0 0.8 0.8 1.56 0. 8 0. 4 Streptococcus aecalis var. Zymogenes 1. 56 3. 12 1. 56 0.8 1. 56 Streptococcus faecalis gr. D 5 434 IP 1. 56 1. 56 1. 56 0.8 1. 56 Streptococcus faecalis gr. D F 262-.- 1. 56 1. 56 3. 12 1. 56 1. 56 Sarcina lutea ATCC 9341 25 50 6. 25 6. 25 12. 5 Listeria monocytogenes 5 844 IP 0. 8 1. 56 0. 4 0. 4 1. 56 Listeria monocytogenes Ramisse type 0.8 1. 56 0. 4 0. 4 0. 8 Listeria monocytooenes Joubert E 102-- 0. 8 1. 56 0. 4 0. 4 0. 8 Erysipelothria: insidiosa E 159 $0. 05 0. 1 0.2 50. 05 $0. 05 Erysipelothriz A 27-. 3. 12 3. 12 0.2 0. 1 50. 05 Bacillus cereus ATCC 10 702 0. 1 0. 1 0. 2 $0. 05 50. 05 Bacillus ccreus AICO 9 634-- $0. 05 0. 2 0.4 $0. 05 0. 1 Bacillus subtilis Leugar A 156 0. 1 0 1 0. 1 50. 05 50. 05 Bacillus subtilis Searle 0. 1 0.1 0.2 $0. 05 50. 05 Pasteurella septica type B 12 0.2 0.8 1. 56 1. 56 50. 05 Pasteurella septica type C 5 621-... 0. 8 0.8 0. 1 50. 05 0.2 Pasteurella pseudotubcrculosis C 114-. 6.25 6. 25 1. 56 1. 56 3. 12 Pasteumlla J oubert E 99 6. 25 5. 25 6. 25 3. 12 6. 25 Escherichia coli A 223 IP..- 1. 56 1. 56 1. 56 0. 8 0. 8 Escherichia coli Monod D 198- 0. 8 1. 56 1. 56 0.8 1. 56 Escherichia coli 111 B4 A... 1. 56 1. 56 3. 12 1. 56 3. 12 Escherichia coli 26 B6 A.- 1. 56 1. 56 3. 12 1. 56 3. 12 Escherichia coli 55 B5 A--- 1. 56 1.56 3. 12 1. 56 3. 12 Escherichia coli ATCO 10 536 1. 56 0. 8 1. 56 0. 8 0. 8 Klebsiella pneumoniae E 155.. 25 12. 5 1. 56 1. 56 12. 5 Aerobacter aerogenes E 115-. 0.8 0.8 12.5 3. 12 3. 12 Citrobacter V1 1. 56 6. 25 3. 12 1. 56 6. 25 Salmonella typhi O 901. 6. 25 6. 25 3. 12 3. 12 6. 25 Salmonella para A A 220 IP.. 12. 5 25 6. 25 6. 25 3. 12 Salmonella para B A 214 IP..- 3. 12 3. 12 1. 56 1. 56 3. 12 Salmonella pullorum gallinarum 465 6. 25 25 3. 12 1. 56 1. 56 Shigella dysenteriae 5 728 IP 1. 56 3. 12 1. 56 1. 56 8. 12 Shigella sonnci D 374..... 1. 56 3. 12 1. 56 0. 8 0. 8 Morazella lwojfi D..-. 50 50 50 50 50 Pseudomonas aeruainisa B 74. 50 50 50 50 50 Bordetella bronchiseptica 25 25 50 TABLE 1-00-ntinued 8 $0.05 0. 2 0. 2 0.05 $0.05 Staphylococcus 209 IP 0. 4 0. 8 1. 56 0. 2 0. 2 Staphylococcus aureus yervais D 319. 0.4 0. 8 1. 56 0. 2 $0. 05 Staphylococcus aureus E 120 0. 2 0. 8 0. 08 50. 05 $0. 05 Staphylococcus albus P C 1 200. 0.1 0. 2 0. 2 50. 05 50. 05 Streptococcus pyogcnes gr. A 561 IP. 1. 56 6. 25 1. 56 0.4 0. 2 Streptococcus pyogeucs gr. A D 313 0. 2 6. 25 0. 8 0.4 50. 05 Stre, s gr. B- 1.56 6.25 1.56 0.4 0. 2 Streptococcus pyogenes gr 0.4 6. 25 1. 56 0. 4 0. 2 Streptococcus faecalis var. Zymogen 1. 56 12. 0. 8 0.4 0. 2 Streptococcus faecalis var. Zymogenes. 1. 56 12. 5 0.8 0. 4 0. 2 Streptococcus faccalis gr. D F 262.. 1. 56 12. 5 1. 56 0. 2 0. 2 Sarcirta lutea ATCC 9341 12. 5 3. 12 50 1. 56 0. 8 Listeria monocytoyeues 5 844 IP 1. 56 1. 56 1. 56 0.4 0. 2 Listeria mouocytoyeues Ramisse type 1. 56 0. 4 0. 2 0. 2 Listeria morwcytogenes Joubert E 102. 0. 4 1. 56 0. 4 0. 2 Erysipelothriz insidiosa E 159 s 0. 05 0. 2 0. 2 0. ()5 Erysipelothriz insidiosa A 27- 50. 05 0. 2 50 05 50. 05 Bacillus ccreus ATCC 702.. 0.4 0. 2 0. 1 50. 05 Bacillus cereus ATCC 9 634.-. 0.4 0. 2 0. 2 $0. 05 Bacillus subtilis Leugar A 156. 0.4 0.8 0.1 50. 05 Bacillus subtilis Searle 0.4 0. 2 50. 05 $0. 05 Pasteurella septica type B 55 129 $0. 05 0.1 50. 05 Pasteurella septica type C 5 621--.- O. 4 0. 4 0. 4 Pasteurella L C 114- 3.12 0. 4 0. 4 Pasteurella J oubert E 99 6. 25 0. 4 0.4 Escherichia coli A 223 IP.- 0. 8 0. 4 0. 2 Escherichia coli Monod D 198-. 0.8 0. 4 0.4 Escherichia coli 111 B4 A- 1. 56 0. 8 0.4 Escherichia coli 26 B6 A.-- 1. 56 0. 4 0.4 Escherichia coli 55 B5 A- 1. 56 .8 0.8 0. 4 Escherichia coli ATCC 10 536 0.8 56 0. 4 0.4 .Klebsiella, E 155 12.5 0.8 3.12 1.56 Aerobacter aerogenes E 1 1. 56 2. 5 0.4 0.4 Citrobac-ter V1 1. 56 12. 5 0.4 0.4 Salmonella typhi O 901 6.25 1. 56 0.4 0.4 Salmonella para A. A 220 IP-. 1... 5 3.12 3. 12 3. 12 Salmonella para B. A 214 IP.... 6. 1. 56 1. 56 0. 4 Salmonella pullorum galliriarum 465- 6. 25 3.12 0.4 0.4 Shigella dysenteriae 5 728 IP. 3.12 0.4 0.4 0. 4 Shigella sonnei D 374 3.12 0. 8 0. 4 0. 2 Morazella lwoffi D 50 25 50 Pseudomonas oerugiuisa B 50 50 50 50 Bordetella bronchiseptica 50 25 50 Staphylococcus lo'adonii 0. 1 0 05 0. 05 0. 05 5o. 05 Staphy1ococcus 209 IP 0.4 0. 4 0. 2 0. 2 0. 2 Staphylococcus aureus Gervais D 319. 0. 4 0. 2 0. 2 0.1 0. 2 Staphylococcus aureus E 120 0.4 50. 05 go. 05 g0. 03 go. 05 Staphylococcus albus PC 1 200-.. 0. 1 $0. 05 $0. 05 50. 05 0. 1 Streptococcus pyogenes gr. A 561 IP.. 0.4 0. 2 0.4 0. 4 0. 2 Streptococcus pyoyeues gr. A D 313 0.8 50. 05 0.4 0.4 0. 1 Stre 7 g1. 0.4 0.2 0.2 0.4 0.2 Streptococcus H gr. 0 0.4 0. 2 0. B 0.4 0. 2 Streptococcus faecalis var. Zymogenes 0.4 0. 2 0.4 0.4 0. 2 Streptococcus faecalis gr. D 5 434 IP. 0. 4 0. 2 0. 2 0. 4 0. 2 Streptococcus faecalis gr. D F 262 0. 8 0.4 0. 4 0.4 0.4 Sarciua lutea ATCC 9341 25 12. 5 3. 12 3. 12 3. 12 Listeria Monocytoyeues 5 844 IP 0.8 0. 2 0.8 0. 2 0. 2 Listeria monocytogenes Ramisse type 0.8 0. 2 0.8 0. 4 0. 2 Listeria monocytogenes Joubert E 102- 0. 8 0. 4 0.8 0.4 0. 2 Erysipelothriz iusidiosa E 159 0.8 50. 05 0. 1 0. 05 0. 1 Erysipelothriz iusidiosa A 27.-. 0. 1 $0. 05 S0. 05 $0. 05 $0. 05 Bacillus cereus ATCC 10 702.. 0. 2 0. 2 O. 1 $0. 05 $0. 05 Bacillus cereus ATCC 9 634.-- 0. 1 0.4 0. 1 50. 05 $0. 05 Bacillus subtilis Leugar A 156 0. 8 0.4 0. 1 50. 05 50. 05 Bacillus subtzlis Semle 0. 4 0. 2 0. 1 50. 05 $0. 05 Pasteurella septica type B 129 50. 05 $0. 05 50. 05 Pasteurella septica type C 5 21 50. 05 0. 05 50. 05 Pasteurella pseudotuberculosis C 114. 0. 1 0. 2 0. 8 Pasteurella J oubert E 99 0. 2 0. 4 1. 56 Escherichia coli A 223 IP.- 0.4 0.2 0.8 Escherichia Monod D 198.. I 0. 4 0.4 1. 56 Escherichia 111 B4 A. 0. 8 0.4 1. 56 Escherichia 26 B6 A--- O. 4 0. 4 1. 56 Escherichia 55 B5 A. 0.4 0.4 1. 56 Escherichia ATCC 10 536..- 0.4 0.4 1. 56 10"" p Elfi'i 0.8 0.8 3.12 Aerobacter aerooertes E 115--. 0.4 0.4 1. 56 Citrobactor Vi 0.4 0. 8 1. 56 Salmonella typhi O 901 0. 4 0. 8 3.12 Salmonella para A A 220 IP 1. 56 1. 56 6. 25 Salmonella para B A 214 IP-... 0.8 0.8 3.12 Salmonella pullorurn yallinarum 465- 0. 4 0. 8 3. 12 Shiyella Dysenteriae 5 728 IP 0. 2 0. 4 1. 56 Shigella sonnei D 374 0.4 0. 4 1. 56 Morazella lwofii D. 6. 25 25 25 Pseudornonas aeruginisa B 74 25 50 50 Bordetella bro'nchiseptica 3. 12 25 25 S s l 0. 1 S0. ()5 0. 05 $0. 05 $0. 05 Staphylococcus 209 IP 0. 4 0. 2 0. 4 0. 2 0. 2 Staphylococcus aureus Gerv 0. 2 0. 2 0. 4 0. 2 0. 2 Staphylococcus aureus E 0. 2 0. 1 0. 1 50. 05 0. 1 Staphylococcus albus PO 1 200.-. 0.1 0. 1 $0. 05 0. 1 0. 1 Streptococcus pyogenes gr. A 561 0.4 0. 4 0.4 O. 4 0. 4 Str "gnADtli 0.4 0.1 0.8 0.2 0.1 Streptococcus gr. 0.4 0. 4 0. 4 0.4 0. 4 Streptococcus pyogeues gr. 0 0. 8 0.4 0. 4 0. 2 0. 4 Streptococcus faecalis var. Zymogenes. 0. 8 0. 4 0. 4 0. 4 0. 4 Streptococcus faecalis gr. D 5 434 IP. 0. 4 0.4 0. 4 0. 4 0.4 Streptococcus faecalis gr. D F 262. 0. 8 0.4 0. 8 0.4 0.4 Sarclna lutea ATCC 9341 3. 12 3. 12 3. 12 3. 12 6. 25

TABLE 1Continued Listeria monocytogenes 5 844 IP 1. 56 0. 4 3. 12 3. 12 3. 12 Listcria monocytogenes Ramisse type IV 1. 56 0. 4 0. 4 0. 4 0.4 Listeria monocytogenes Joubert E 102. 1. 56 0. 4 0. 2 0. 4 0. 4 Erysipelalhrir Insidiosa E 159 0. 2 0. 1 0.2 0. 1 0. 1 Erysipelothria: Insidiosa A 27- 0. 2 S- 1 0- 1 0. Bacillus cereus AICO 702. 0. 2 $0 05 0. 2 $0. 05 50. 05 Bacillus cereus ATCC 9 634. 0. 2 0 05 0. 1 50. 05 0. 1 Bacillus subtilis' Leugar A 156- 0. 2 0. 1 0. 1 50. 05 $0. 05 Bacillus snbiili: Searle- 0. 2 s0 05 0. 1 0. 05 50. 05 Pasteurella Septica type B 55 129 1. 56 1 56 0. 4 0. 4 0. 8 Pasteurella Septica type C 5 621 0. 1 0. 1 $0. 05 $0. 05 50 05 Pasteurella pscurlctuberculusi: C 114. 0.4 0.4 0. 4 0. 4 1 0.8 Pasteurella Joubert; E 99- 3. 12 0. 8 0. 8 1. 56 1. 56 Escherichia coli A 223 IP 3. 12 0. 8 0. 4 0. 4 3. 12 Escherichia coli Monod D 3. 12 0. 8 0. 8 0. 8 3. 12 Escherichia coli 111 B4 A. 3. 12 1. 56 1. 56 1. 56 3. 12 Escherichia C017 26 B6 A. 3. 12 1. 56 1. 56 0. 8 3. 12 Escherichia coli 55 B5 A 3. 12 0.8 1. 56 0. 8 3. 12 Escherichia coli ATCC 10 536 3- 12 0 8 55 55 12 K'lcbaiclla f."- E 155 6. 1. 56 1. 56 0. 4 6. 25 Acrobacter azrogenu E 115 3. 11 0- 3 3 8 3- 12 Citrobactgr Vi 3. 12 1. 56 1. 56 1. 56 3. 12 Salmonella typhi O 901 3. 12 1. 56 1. 56 1. 56 3. 12 Salmonella para A A 220 IP. 6. 25 6. 25 3. 12 3. 12 12. 5 Salmonella para B A 214 IP 3. 1 1. 56 12 3 12 Salmonella pulloru'm gallinaram 465. 3. 12 1. 56 0- 8 0. 4 3. 12 Shiqella dyunteriac 5 728 IP 1. 56 0. 4 0. 5 0. 4 3. 12 Shiaella zonnei D 374---- 1. 56 0. 4 0. 8 0.8 3. 12 Morarella lwojfi D 5 59 25 59 59 Pszudo'mona: aeruainis 50 50 50 50 50 Bordetella bronchissptica 25 25 25 25 25 TABLE 2.ANTIBACTERIAL ACTION IN VITRO AGAINST 25 TABLE 3, NTIF 3 [TY I I MYOOBACTERIUM TUBERCULOSIS H 37 RV STRAIN A N v TRO [Minimum inhibiting concentrations in mcg.lm1.] [Mlmmum mhlbmng concentratmns in mcg'lml'] Inoculated with 0.1 mg., Inoculated w1th 0.01 mg., 10290 10289 10291 10385 10337 10453 y) ay) Pr d c 2 18th Trich. mentagrophytes t g 6th 1 4 30 30 30 30 30 30 ric men 70 ea 3.72 3.75 will.

7 5 Trich. 'menta r0 h tes 05 5 (mice) 0.1 0. 75 Epiderm. flocossnm E24- 10 10 10 1. 25 1. 5 5 Oryptococcus noformans 25 25 5 75 5 5 3 Epiderm. flocossum 193 1. 25 10 10 0. 25 1. 25 1. 2 IP 20 5 5 2. 5 5 5 5 5 Candida albican: 200 IP. 10 5 0 0 75 5 Candida albicans No. 2.-.. $0. 5 10 10 10 2.5 5 5 Candida tropicalis No. 1 0. 25 0. 75 1. 25 0. 15 0.15 0. 25 Candida pseudotwpimlis 0. 75 0. 75 2. 5 0. 15 0. 75 1- 25 0. 75 1. 25 1. 25 0. 25 0. 75 0. 75 c did M o. 75 1. 25 1. 25 1. 25 2. 6 2 5 40 Candida para-krusei 2. 5 2. 5 2. 5 0. 25 0. 75 0. 75 Microspo um gypgguqn 2. 5 5 5 0. 75 1. 25 2. 5 0. 75 0. 75 0. 75 0. 25 0. 75 0. 75 0. 1.222 1. 25 0. 75 0. 72 0.272 1. 2. 5 0. 5 1. 2 1 g 1 25 0. 75 0 g 10452 10383 10384 10743 10744 10137 2. 2. 5 0. 75 1. 2 0. 75 1.25 1. 25 0. 25 o. 75 0. 75 mmtwophvm 1. 25 1. 25 1. 25 0.15 0.15 0.15 20 5 0. 75 0. 75 o. 75 o. 25 0. 75 0. 75 fl fl fl 0. 75 0. 75 0. 75 o. 75 o. 75 0. 75 Q 51 0. 75 0. 75 0. 75 o. 75 o. 75 0. 75 'ko. 1 0. 25 0.25 0. 05 0.15 0.15 0.05 15 g 05 005 Q1 Epiderm flocosswm E24 30 20 30 0. 75 0. 75 1. 25 o. 75 o. 75 0. 75 Gil/1110mm "ofoimanh 0. 75 0. 75 1. 25 0. 25 o. 75 0. 75 1 193 1. 25 2. 5 2. 5 go. 05 0.15 1. 25 25 5 5 2. 5 10 10 5 5 10 gand da allz cans i100 1219. 30 2.5 5 30 30 30 s o. j g g 2 2 Candida tropicalis No. 0. 75 o. 75 o 75 0125 0. 75 o. 75 0471414 pmdoimpiml" 1.25 5 5 0.75 1.25 1.25 2 2.5 2.5 2.5 0.75 2.5 2.5 1.25 5 5 0. 75 1.25 2.5 10 1o 10 0. 75 0 75 g 9111mm" 0. 75 r 3.72 S372 "0. 0.15 0. 25 0. 75 0. 1 0 25 0. 25 15 0, 75 7 1 1 025 0, 75 10933 10934 11064 11071 10829 10827 2. 5 2. 5 2. 5 1. 25 2. 5 2. 5

2. 5 5 5 0. 75 0. 75 5 Trich. Montagrophytcs 2. 5 5 10 2. 5 2. 5 5 403 20 1. 25 2. 5 5 2. 5

5 5 5 0. 25 0.25 0. 25 Trich. Mentagrophytes 1. 25 1. 25 1. 25 0. 25 0. 75 0- 75 25 2. 5 10 5 10 10 0. 75 1. 25 5 Trich. meniagrophytes 2. 5 2. 5 2. 5 0. 75 0. 75 0. 75 mic 1. 25 2. 5 10 0. 25 0. 75 0. 75 0. O5 0. 1 0. 15 Epiderm. floccssu'm E 0. 75 0. 75 1. 25 0. 15 0. 75 0. 75 24 10 30 10 5 10 5 1. 25 2. 5 2. 5 0. 75 1. 25 1. 25 Ory'ptococcus 0. 1 0. 25 0. 75 50. 05 0. 25 0. 75 noformans 10 20 0. 25 0.25 0. 75 50. 05 0. 15 0. 15 Epiderm. flocossnm 193 0. 75 0. 75 0. 75 0. 75 0. 75 0. 75 IP 5 2. 5 0. 25 0. 25 0. 75 0. 1 0. 25 0. 75 Candida a icans 200 I1- 2. 5 s0. 5 5 5 10 10 5 5 5 Candida albicans N0. 2 5 5 0. 75 0. 75 0. 75 0. 15 0. 15 0. 15 Candida tropicalis 0. 25 2. 5 2. 5 0. 1 0. 0. 75 N0. 1 10 20 2. 5 2. 5 5 1. 25 2. 5 2. 5 Candida 2. 5 2. 5 2. 5 0. 25 2. 5 2. 5 pseudoirapicalis 2 1. 25 1. 25 2. 5 2. 5 2. 5 0.25 0. 75 1. 25 Candida krnsei 5 10 2. 5 2. 5 2. 5 2. 5 2. 5 2. 5 Candida para-Masai"-.- 20 2. 5 5 5 2. 5 2. 5 2. 5 crosp azlp 5 5 5 1, 25 2, 5 5 D 5 10 10 10 5 10 10 75 Microsporam canis E 25.. 30

TABLE 3C0ntinued TABLE 4.ANTIPARASITICIDAL AGT1'V1TY I N vivo 10946 10746 11141 11142 11025 11026 P r entage (lepalasitisation Syphacia Syphacia 20 30 2 5 2.5 5 20 Obvzlam Product Obvelaw Trich.menta To 11 tes 1.25 5 10 30 10 Trich. mentagrophytes 44 (mic 10 5 10 30 Epiderm. flocossum E 0 Cryptococcus 2 neoformans 30 1.25 50.5 20 10 g Epider'm. flocomim 1931p 1. 25 1. 25 30 Candida albz'cans 200 IP 1.25 30 1. 25 50.5 1. 25 1.25

Candida albicans 2 No.2 30 30 30 25 20 30 3% Candida tropicalis N0. 1 30 30 30 30 30 30 Candida 37 pseudotropicalis 2- 5 30 1. 25 0.5 1. 25 2.5 37 Candida krusei 30 30 30 30 30 30 37 ggndida para-mm.-." 30 30 25 10 30 20 icros 0mm seam D"? 2.5 2 5 10 25 Microsporum canis E 25 5 5 25 25 20 Nipposirongylus mans Hymenolepszs nana 11245 11246 11262 11263 11273 Stronglus Young cy fl Product larvae adults Adults colds Adults Trich. mentagrophytes 40 5 20 5 5 Trich. mentagroplwtes G 5 10 10 Trich. mentagrophytes mic 5 30 10 10 Epiderm. flocossum 2 5 5 1. 25 5 Cryplococcus nofermans. 10 30 1. 25 O. 5 Epiderm. flocossum 193 IP 5 5 5 5 3 2202 12s .0 r

("I 1 aa 16117718 0.

Candida tropicalis No.1- 10 30 30 30 25 TABLE URINARY EXCRETION OF THE RAT Candida [Oral administration] paeudoiropicalis 2- 0.5 1. 25 5 1. 25 5 Candida krusei. 30 30 30 Active bacteriological maten'al in the urine- Candida para-mum"..- 2.5 30 30 30 25 Time ft administration Microsporum gypseu'm 1) 5 30 25 10 20 Dose, 0-1 1-2 -24 4-6 0-3 3-24 11416108210114?" cam-9 E 5 5 1O 20 Product mgJkg. hr. hrs. hrs. hrs. hrs. hrs.

10743 100 Mcg./m1.urine 1.5 1.3 0 O 0.29

Mcg.total 1.2 1.56 0 0 0.17

40 10744 100 McgJmLurine" 0.1 0 0 0 0 Meg. total 0.3 0 0 0 0 10933"--. 100 Mcg./m1.urine 0 O 0 0 Meg. total 0 0 0 0 10932"--- 100 Mcg./m1.urine 0 0.34 0.12 0.15 0. Meg. total 0 0.37 0.23 0 39 0.

11070 100 Mcg./m1.urine 0.35 0.27 0.23 0.8 0.38 0. 075 Meg. total 5.7 0.82 0.84 3.92 2.05 0.98

10452 100 Mcg./m1.urine 0 0.28 2.6 3.4 0 0 Meg. total 0 1.85 3.12 3. 74 0 0 10683".-. 100 McgJmLurine 0 0.58 4.1 0 0 0 Meg. total 0 0.46 1.5 0 0 0 TABLE 6.-INTERNAL ANTISEPTIC ACTION Modification of the Flora of Healthy Animals 2 Percentage variation in faeeal germs Dose inmgJkg. Before 3 days body- Dilution adminis- After After After after Product weight of faeces tration 1 day 352.375 7 days treatment 10": 1g 38. g 50 80 10 2 2 20 10452 10+ 100 30 10 10 s5 10- 100 0 0 0 100 10": 100 50 12 56 0' 100 45 10 40 10633 10- 100 0 0 0 30 10 100 0 0 0 100 100 g2 g5 92 76 r 00 0 5 80 92 mm 10- 70 70 100 100 10' 100 100 13 25.2 20 42 100 30 0 50 10746 10- 100 0 10 20 50 10 0 0 0 0 0 10: 100 10g 100 100 100 0 100 50 90 45 11139 10- 100 8 100 100 5o 10 0 0 0 0 0 is: 180 100 lgO 100 100 0 100 0 40 37 11140 10 100 30 100 40 20 10 100 25 25 60 60 TABLE 6C0ntinued Percentage variation in faecal germs Dose mmg./kg. Before 3 days body- Dilution adminis- Alter After After after Product weight of faeces tration 1 day 3 days 7 days treatment 18? 188 138 P2 100 100 11052 100 0 1o 70 35 10- 100 0 5 70 100 10": 180 100 100 100 0 0 100 100 80 10 100 50 50 50 10 100 10 50 50 50 10-2 100 103 lgl) 100 100 0* 00 0 0 50 100 11262 10- 100 so 40 10 100 10 100 40 10 0 100 10': 100 100 100 120 100 10 00 100 00 0 100 11263 10- 100 70 so 0 40 10' 100 60 0 10 1 Given daily for 7 days.

TABLE 9 TABLE 7.INTESTINAL ANTISEPTIC ACTION [Animals infected with S. typhimurium] Dose in Ins/kgbody- Percentage vanation ll! faceal germs weight given Dilu- Before 3 days daily tion adminafter Prodfor 7 of istra- After After After treatuct days faeces tion 1 day 3 days 7 days ment TABLE 8.TOXICITY IN MICE LDu LDsn Intraperi- Intraperl- Product toneal Oral toneal Oral 11140 1 g./kg

Compounds 12004 and 12003 were subjected to a search of bacteriostatic properties in vitro.

For this purpose a series of dilutions to /2 was effected in agar medium, the contents of active compound in the culture medium ranging from 0.05 to 50 micrograms (meg) per ml. Various microbial strains, gram+ and gram as well, were used to inoculate the medium. After 24 hours in an oven at 37 C., the minimum inhibiting concentrations (M.I.C.) were determined.

Youmans medium was employed to study tuberculostatic activity, readings were taken on the sixth, twelth and eighteenth day after inoculation of the medium with Mycobacterium tuberculosis (H37RV strain).

Table 9 shows the results obtained after 24 hours in screening tests. In the conditions of the experiment, the bacteriostatic activity of compound 12004 towards gram+ germs as well as gram" germs was medium-sized.

Gram+ germs:

Staphylococcus lona'onii 1.5;6 Staphylococcus 209 IP 1.56 Staphylococcus aureus Gervais D 319 3.12 Staphylococcus aureus E 3.12 Staphylococcus albus P C I 1 200 0.8 Streptococcus pyogenes gr. A 561 IP 12.5 Streptococuus pyogenes gr. A D 313 12.5 Streptococcus agalactiae gr. B 6.25 Streptococcus pyogenes gr. C 6.25 Streptococcus faecalis var. Zymogenes 12.5 Streptococcus faecalis gr. D 5 434 IP 12.5 Streptococcus faecalis gr. D F 262 12.5 Sarcina lutea ATCC 9341 50 Listeria monocytogenes 5 844 IP 12.5 Listeria monocytogenes Ramisse type IV 12.5 Listeria monocytogenes Joubert E 102 12.5 Erysipelothrix insidiosa E 159 6.25 Erysipelothrix insidiosa A 27 12.5 Bacillus cereus ATCC 10 702 1.56 Bacillus cereus ATCC 9 634 1.56 Bacillus subtilis Leugar A 156 1.56 Bacillus subtilis Searle 1.56 Pasteurella septica type B 55 129 3.12 Pasteurella septica type C 5 621 3.12 Pasteurella pseudotuberculosis C 114 12.5 Pasteurella Joubert E 99 12.5 Escherichia coli A 223 IP 12.5 Escherichia coli Monod D 198 6.25 Escherichia coli 111 B4 A 12.5 Escherichia coli 26 B6 A 12.5 Escherichia coli 55 B5 A 12.5 Escherichia coli ATCC 10 536 6.25 Klebsiella pneumoniae E Q 12.5 Aerobacter aerogenes E 115 25 Citrobacter vi 50 Salmonella typhy O 901 50 Salmonella para A A 220 IP 50 Salmonella para B A 214 IP 50 Salmonella pullorum gallinarum 465 50 Shigella dysenteriae 5 728 IP 12.5 Shigella sonnei D 374 12.5 Moraxella lwojfi D 50 Pseudomonas aeroginosa B 74 50 Bordetella brunchiseptica 50 As to compound 12003, the results appear in Table 10. 

